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access arraytm ifc  (fluidigm)


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    fluidigm access arraytm ifc
    Access Arraytm Ifc, supplied by fluidigm, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ifc+controller+for+access+arraytm+system/Control+Line+Fluid+Kit/us10501786-1624-65-69
    Average 92 stars, based on 17 article reviews
    access arraytm ifc - by Bioz Stars, 2026-11
    92/100 stars

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    Related Articles

    Control:

    Article Title: Nucleic acid encoding reactions
    Article Snippet: .. Preparing Amplicons The following documents may be consulted as references for this protocol: Fluidigm® IFC Controller for ACCESS ARRAYTM System User Guide (PN 68000157); Fluidigm® Control Line Fluid Loading Procedure Quick Reference (PN68000132); and Agilent DNA 1000 Kit Guide. .. The following Reagents were used for this protocol and were stored at −20° C.: FastStart High Fidelity PCR System, dNTPack (Roche, PN 04-738-292-001); 20× ACCESS ARRAYTM Loading Reagent (Fluidigm, PN 100-0883); 1× ACCESS ARRAYTM Harvest Solution (Fluidigm, PN 100-1031); ACCESS ARRAYTM Barcode Library for Illumina Sequencers—384 (Bidirectional) (Fluidigm, PN 100-3771); target-specific primer pairs tagged with universal tags (CS1 forward tag, CS2 reverse tag), including 50 μM CS1-Tagged TS Forward Primer and 50 μM CS2-Tagged TS Reverse Primer; and template DNA at 50 ng/μL. (The 1× ACCESS ARRAYTM Harvest Solution (Fluidigm, PN 100-1031) is not packaged for individual sale.

    Article Title: Multiplexed CRISPR-based microfluidic platform for clinical testing of respiratory viruses and identification of SARS-CoV-2 variants
    Article Snippet: .. Loading the control line fluid (single control line fluid syringe in Fluidigm Kit) a. ..

    Article Title: Nucleic acid encoding reactions
    Article Snippet: Other reagents were stored at room temperature, including PCR Certified Water (Teknova, PN W330); DNA Suspension Buffer (10 mM Tris HCl, 0.1 mM EDTA, pH8.0) (Teknova, PN T0221); and Agilent DNA 1000 Chips (included in the Agilent DNA 1000 DNA kit) (Agilent). .. The following equipment and consumables were used for this protocol: 1.5 mL or 2 mL microcentrifuge tubes; Microcentrifuge with rotor for 2 mL tubes; Microcentrifuge with rotor for 0.2 mL PCR tube strips; Centrifuge with plate carriers; Agilent 2100 BioAnalyzer (Agilent); 96-Well Reaction Plate; MicroAmp Clear Adhesive Film (Applied Biosystems, PN 4306311); IFC Controller AX (2 quantity, pre- and post-PCR) (Fluidigm); FC1 Cycler (Fluidigm); 48.48 ACCESS ARRAYTM IFC s (Fluidigm); and Control Line Fluid Syringes (Fluidigm, PN 89000020). .. Multiplex PCR on the ACCESS ARRAYTM IFC was performed according to the instructions as detailed in Chapter 6—Multiplex PCR on the 48.48 ACCESS ARRAYTM IFC of the Fluidigm ACCESS ARRAYTM System for Illumina Platform User Guide.

    Article Title: Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies
    Article Snippet: .. Before the first use of each IFC, the chips were primed with the Control Line Fluid Kit in Juno System (Fluidigm, United States) according to manufacturer’s guidelines. ..

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: 48.48 Dynamic Array IFC for Gene Expression , Fluidigm , SKU BMK-M-48.48. .. Control Line Fluid Kit—48.48 , Fluidigm , Cat# 89000020. .. 20X GE Sample Loading Reagent , Fluidigm , Cat #100-7610.

    Article Title: Methods and kits for extracting nucleic acids from paraffin embedded samples
    Article Snippet: FIG. 9C shows that the number of targets identified by the Fluidigm kit had about a 45% improvement, which was operator independent. .. Thus, the Fluidigm® test kit passed the quality control analysis according to all three criteria. .. When assessed by criteria (2) and (3), results were improved to an extent that would substantially enhance expression analysis for purposes of research and/or clinical diagnosis.

    Article Title: Downregulation of CSF-derived miRNAs miR-142-3p and miR-17-5p may be associated with post-dural puncture headache in pregnant women upon spinal anesthesia
    Article Snippet: Fluidigm’s microfluidic gene expression system was used for RT-qPCR analysis. .. Dynamic Array (96.96) integrated fluidic circuit (IFC) (Fluidigm, South San Francisco, CA) was injected with Control Line Fluid Kit and primed using the IFC Controller HX (Fluidigm). .. Mixtures for assay inlets and sample inlets were prepared using miScript Microfluidics PCR Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Polymerase Chain Reaction:

    Article Title: Nucleic acid encoding reactions
    Article Snippet: Other reagents were stored at room temperature, including PCR Certified Water (Teknova, PN W330); DNA Suspension Buffer (10 mM Tris HCl, 0.1 mM EDTA, pH8.0) (Teknova, PN T0221); and Agilent DNA 1000 Chips (included in the Agilent DNA 1000 DNA kit) (Agilent). .. The following equipment and consumables were used for this protocol: 1.5 mL or 2 mL microcentrifuge tubes; Microcentrifuge with rotor for 2 mL tubes; Microcentrifuge with rotor for 0.2 mL PCR tube strips; Centrifuge with plate carriers; Agilent 2100 BioAnalyzer (Agilent); 96-Well Reaction Plate; MicroAmp Clear Adhesive Film (Applied Biosystems, PN 4306311); IFC Controller AX (2 quantity, pre- and post-PCR) (Fluidigm); FC1 Cycler (Fluidigm); 48.48 ACCESS ARRAYTM IFC s (Fluidigm); and Control Line Fluid Syringes (Fluidigm, PN 89000020). .. Multiplex PCR on the ACCESS ARRAYTM IFC was performed according to the instructions as detailed in Chapter 6—Multiplex PCR on the 48.48 ACCESS ARRAYTM IFC of the Fluidigm ACCESS ARRAYTM System for Illumina Platform User Guide.

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Adhesive:

    Article Title: Nucleic acid encoding reactions
    Article Snippet: Other reagents were stored at room temperature, including PCR Certified Water (Teknova, PN W330); DNA Suspension Buffer (10 mM Tris HCl, 0.1 mM EDTA, pH8.0) (Teknova, PN T0221); and Agilent DNA 1000 Chips (included in the Agilent DNA 1000 DNA kit) (Agilent). .. The following equipment and consumables were used for this protocol: 1.5 mL or 2 mL microcentrifuge tubes; Microcentrifuge with rotor for 2 mL tubes; Microcentrifuge with rotor for 0.2 mL PCR tube strips; Centrifuge with plate carriers; Agilent 2100 BioAnalyzer (Agilent); 96-Well Reaction Plate; MicroAmp Clear Adhesive Film (Applied Biosystems, PN 4306311); IFC Controller AX (2 quantity, pre- and post-PCR) (Fluidigm); FC1 Cycler (Fluidigm); 48.48 ACCESS ARRAYTM IFC s (Fluidigm); and Control Line Fluid Syringes (Fluidigm, PN 89000020). .. Multiplex PCR on the ACCESS ARRAYTM IFC was performed according to the instructions as detailed in Chapter 6—Multiplex PCR on the 48.48 ACCESS ARRAYTM IFC of the Fluidigm ACCESS ARRAYTM System for Illumina Platform User Guide.

    Injection:

    Article Title: Downregulation of CSF-derived miRNAs miR-142-3p and miR-17-5p may be associated with post-dural puncture headache in pregnant women upon spinal anesthesia
    Article Snippet: Fluidigm’s microfluidic gene expression system was used for RT-qPCR analysis. .. Dynamic Array (96.96) integrated fluidic circuit (IFC) (Fluidigm, South San Francisco, CA) was injected with Control Line Fluid Kit and primed using the IFC Controller HX (Fluidigm). .. Mixtures for assay inlets and sample inlets were prepared using miScript Microfluidics PCR Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

    Single Cell:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    DNA Library Preparation:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Quantitative RT-PCR:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Gene Expression:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    RNA Sequencing:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Mutagenesis:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Knock-Out:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Recombinant:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS

    Software:

    Article Title: The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition
    Article Snippet: Reagent or resource Source Identifier Rat anti- Gr1 (Ly6G) APC-Cy7, Clone RB6-8C5 BD Biosciences Cat# 557661; RRID:AB_396775 Rat anti-mouse c-Kit PE, Clone 2B8 BD Biosciences Cat# 553355; RRID:AB_394806 Rat anti-mouse c-Kit PE-Cy7, Clone 2B8 BD Biosciences Cat# 558163; RRID:AB_647250 Rat anti-mouse Ly-6A/E (Sca1) PE-Cy7, Clone D7 eBioscience Cat# 25-5981-82; RRID:AB_469669 Rat anti-mouse Ly-6A/E (Sca1) FITC, Clone E13-161.7 BioLegend Cat# 122505; RRID:AB_756190 Streptavidin PE Thermo Fisher Cat# 12-4317-87 Rat anti-mouse Ter119 APC-ef780, Clone TER-119 eBioscience Cat# 47-5921-80; RRID:AB_1548794 Rat anti-mouse Ter119 PE-Cy7, Clone TER-119 BD Biosciences Cat# 557853; RRID:AB_396898 Rat anti-mouse VE-Cadherin ef660, Clone eBioBV13 eBioscience Cat# 50-1441-82; RRID:AB_11219483 Bacterial and virus strains Biological samples Chemicals, peptides, and recombinant proteins 7-aminoactinomycin D (7-AAD) ThermoFisher Scientific Cat# A1310 eBioscience fixable viability dye eFluor 780 eBioscience Cat# 65-0865-14 Hoechst 33258 Merck Cat# 861405 Benzyl alcohol Merck Cat# 108006 Benzyl benzoate Merck Cat# B6630 Mouse SCF PeproTech Cat# 250-03-100ug Mouse FLT3L PeproTech Cat#250-39L-100ug Mouse IL-3 PeproTech Cat#213-13-100ug Hyclone FCS Fisher Scientific Cat# SH30070.03HI Collagenase I Merck Cat# C0130 Hot StartTaq Master Mix QIAGEN Cat# 203445 Iscove’s Modified Dulbecco’s Medium (IMDM) ThermoFisher Scientific Cat# 12440053 Dimethyloxalylglycine (DMOG) Merck Cat# D3695 DAPT Merck Cat# 565770 Dofetilide Merck Cat# PZ0016 RNaseOUT Recombinant Ribonuclease Inhibitor ThermoFisher Scientific Cat# 10777019 SUPERasedIn RNase Inhibitor (20 U/mL) ThermoFisher Scientific Cat # AM2694 BD Pharmlyse BD Biosciences RRID:AB_2869057 ERCC RNA Spike-In Mix ThermoFisher Scientific Cat #4456740 SMARTScribe Reverse Transcriptase Takara Cat #639538 SeqAmp DNA Polymerase Takara Cat #638509 AMPure XP Beckman Coulter Cat # A63881 dNTP set (100 mM) ThermoFisher Scientific Cat# 10297018 Magnesium chloride solution Merck Cat# M1028 Critical commercial assays Taqman assay: Atp5a1 ThermoFisher Scientific Mm00431960_m1 Taqman assay: Dll4 ThermoFisher Scientific Mm00444619_m1 (Continued on next page) e2 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Dead Cell Removal Kit Miltenyi Biotec Cat# 130-090-101 Chromium Next GEM Single Cell 30 v3.1 Kit 10x Genomics Cat# 1000269 NextSeq 500/550 High Output Kit v2 (150 cycles) Illumina Cat# FC-404-2002 Nextera XT DNA Library Preparation Kit (96 samples) Illumina Cat# FC-131-1096 Nextera XT Index Kit v2 Set A (96 indexes, 384 samples) Illumina Cat # FC-131-2001 Nextera XT Index Kit v2 Set D (96 indexes, 384 samples) Illumina Cat # FC-131-2004 CellsDirect One-Step qRT-PCR Kit ThermoFisher Scientific Cat# 11753100 TaqMan Universal PCR Master Mix ThermoFisher Scientific Cat # 4304437 48.48 Dynamic Array IFC for Gene Expression Fluidigm SKU BMK-M-48.48 Control Line Fluid Kit—48.48 Fluidigm Cat# 89000020 20X GE Sample Loading Reagent Fluidigm Cat #100-7610 2X Assay Loading Reagent Fluidigm Cat # 100-7611 Deposited data Smart-Seq2 single cell RNA sequencing data of cells undergoing EHT in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-8362 10x single cell RNA sequencing data of whole PAS in wild type and Ncx1 / mutant embryos This paper ArrayExpress:E-MTAB-10945 Experimental models: Cell lines OP9 Gift from A. Medvinsky N/A OP9-Jag1 Gift from Ana Bigas (Van de Walle et al., 2011) Experimental models: Organisms/strains Mouse: Ncx1 knockout Gift from Simon J. Conway (Koushik et al., 2001) Mouse: 23GFP Our laboratory (Bee et al., 2010; Swiers et al., 2013) Oligonucleotides LacZL: GAC GTC TCG TTG CTG CAT AA IDT (Koushik et al., 2001) LacZR: CAG CAG CAG ACC ATT TTC AA IDT (Koushik et al., 2001) NCXSense: TGA TGA CCG GAG CTG GCA AC IDT (Koushik et al., 2001) NCXAntisense:AGA TCA CAG TCC CTT CCG TG IDT (Koushik et al., 2001) NeoInsert: CAG CGC ATC GCC TTC TAT CG IDT (Koushik et al., 2001) GFP1: GAC GTG AAC GGC CAC AAG TTC A IDT (Bee et al., 2010) GFP2: GTG GCG GAT CTT GAA GTT CAC C IDT (Bee et al., 2010) Oligo-dt30VN (Custom made) IDT N/A Recombinant DNA Software and algorithms Imaris Bitplane RRID:SCR_007370 Microsoft Excel Microsoft RRID:SCR_016137 GraphPad Prism GraphPad Software RRID:SCR_002798 FlowJo BD RRID:SCR_008520 Photoshop Adobe RRID:SCR_014199 (Continued on next page) e4 Cell Reports 37, 110103, December 14, 2021 Article ll OPEN ACCESS .. Reagent or resource Source Identifier Illustrator Adobe RRID:SCR_010279 Zeiss Zen Zeiss https://www.zeiss.com/microscopy/ int/products/microscopesoftware/zen.html R (R-3.2.3 – R-3.6.2) The R Foundation https://www.r-project.org FastQC (Wingett and Andrews, 2018) N/A Cutadapt (Martin, 2011) N/A STAR (Dobin et al., 2013) N/A Seurat (Butler et al., 2018; Hao et al., 2021; Satija et al., 2015) N/A ToppGene (Chen et al., 2009) https://toppgene.cchmc.org/ GSEA (v 4.0.3) (Subramanian et al., 2005) https://www.gsea-msigdb.org/gsea/ index.jsp Louvain (Blondel et al., 2008) N/A Python https://www.python.org/ RRID:SCR_008394 SCANPY (Wolf et al., 2018) N/A G:Profiler (Reimand et al., 2007) N/A SINGuLAR Fluidigm https://www.fluidigm.com/products- services/software/singularanalysis-toolset QoRTs (Hartley and Mullikin, 2015) N/A KEGG (Kanehisa et al., 2017) N/A SCTransform (Hafemeister and Satija, 2019) N/A Canonical Correlation Analysis (CCA) (Stuart et al., 2019) N/A CellRanger (v5.0.0) 10x Genomics https://support.10xgenomics.com DoubletFinder (McGinnis et al., 2019) N/A Other Echo 525 Acoustic Liquid Handler Beckman Coulter https://www.beckman.com/liquid- handlers/echo-525 Membrane Filter, Pores 0.8 mm, 25 mm Diameter Merck Cat# AAWP02500 Article ll OPEN ACCESS



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    fluidigm ifc controller for access arraytm system
    Ifc Controller For Access Arraytm System, supplied by fluidigm, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ifc+controller+for+access+arraytm+system/Flex+Six+Gene+Expression+IFC/us09074204-1606-2-1
    Average 90 stars, based on 1 article reviews
    ifc controller for access arraytm system - by Bioz Stars, 2026-11
    90/100 stars
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