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a2153 10g ampicillin  (Thermo Fisher)


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    Thermo Fisher a2153 10g ampicillin
    A2153 10g Ampicillin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/a2153+10g+ampicillin/Ampicillin/pm34450043-148-63-67
    Average 99 stars, based on 1 article reviews
    a2153 10g ampicillin - by Bioz Stars, 2026-10
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    Affinity Column:

    Article Title: Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains BL21-CodonPlus (DE3)-RIL Competent cell Agilent Technologies Cat# 230245 Rosetta (DE3)pLysS Competent cell Novagen Cat# 70956 One Shot PIR1 Chemically Competent cell Thermo fisher scientific Cat# C101010 Stellar Competent Cell Takara Bio Cat# 636763 Chemicals, peptides, and recombinant proteins Terrific Broth (TB) Thermo fisher scientific Cat# BP97285 isopropyl b-D-thiogalactopyranoside (IPTG) Thermo fisher scientific Cat# R0393 Tris-Base Thermo fisher scientific Cat# BP152-1 HEPES Thermo fisher scientific Cat# BP310-1 Sodium chloride Thermo fisher scientific Cat# S271-10 Glycerol Thermo fisher scientific Cat# G33-500 2-Mercaptoethanol Thermo fisher scientific Cat# O3446I-100 Magnesium chloride Thermo fisher scientific Cat# M33-500 PMSF Sigma-Aldrich Cat# P7626 Imidazole Sigma-Aldrich Cat# I202-2KG DTT Thermo fisher scientific Cat# AAJ1539722 EDTA Thermo fisher scientific Cat# AM9261 Potassium Chloride Thermo fisher scientific Cat# P217-500 Magnesium acetate tetrahydrate Millipore Sigma Cat# M5661-250G Agarose Thermo fisher scientific Cat# BP1356-500 SYBR safe DNA Gel Stain Thermo fisher scientific Cat# S33102 Tobacco Etch Virus (TEV) protease Homemade N/A FspI New England Biolabs Cat# R0135L HindIII-HF New England Biolabs Cat# R3104S BseRI New England Biolabs Cat# R0581L BsaI New England Biolabs Cat# R3733S Proteinase K Thermo fisher scientific Cat# EO0491 Benzonase Nuclease Millipore Sigma Cat# E1014-25KU ATP Thermo fisher scientific Cat# BP413-25 Phusion Hot Start II High-Fidelity PCR Master Mix Thermo fisher scientific Cat# F565L Phusion High-Fidelity PCR Master Mix with HF Buffer Thermo fisher scientific Cat# F531L PrimeSTAR Max Premix Takara Bio Cat# R045A In-Fusion HD Cloning Kit Takara Bio Cat# 639650 Q5 Site-Directed Mutagenesis Kit New England Biolabs Cat# E0554 HiScribe T7 Quick High Yield RNA Synthesis Kit New England Biolabs Cat# E2050S QIAprep Spin Miniprep Kit QIAGEN Cat# 27106 MinElute PCR Purification Kit QIAGEN Cat# 28006 ZymoPURE II Plasmid Gigaprep Kit Zymo Research Cat# D4204-B Mini-PROTEAN TGX Precast Gels Bio-rad Cat# 4561086 Amicon Ultra-15 Centrifugal Filter Units 3kDa NMWL Millipore sigma Cat# UFC900324 (Continued on next page) Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Amicon Ultra-15 Centrifugal Filter Units 10kDa NMWL Millipore sigma Cat# UFC901024 Amicon Ultra-15 Centrifugal Filter Units 30kDa NMWL Millipore sigma Cat# UFC903024 Amicon Ultra-15 Centrifugal Filter Units 50kDa NMWL Millipore sigma Cat# UFC905024 NuPAGE LDS Sample Buffer (4X) Thermo fisher scientific Cat# NP0007 Gel Loading Dye, Purple (6X) New England Biolabs Cat# B7024S Bovine Serum Albumin Sigma-Aldrich Cat# A2153-10G Ampicillin, sodium salt Thermo fisher scientific Cat# BP1760-25 Chloramphenicol, 98%, ACROS Organics Thermo fisher scientific Cat# AC227920250 Kanamycin Monosulfate, USP Grade Gold Biotechnology Cat# K-120-5 SnakeSkin Dialysis Tubing, 7K MWCO, 22 mm Thermo fisher scientific Cat# 68700 Diethyl pyrocarbonate Millipore sigma Cat# D5758-25ML Critical commercial assays Ni-NTA agarose resin QIAGEN Cat# 30430 HiTrap Heparin HP affinity column Cytiva Cat# 17040701 RESOURCE Q anion exchange column Cytiva Cat# 17117701 Superdex 200 10/300 column Cytiva Cat# 28990944 InstantBlue Coomassie Protein Stain Abacam Cat# ISB1L Deposited data Cas12k–sgRNA–target DNA coordinate This study PDB: 7N3P Cas12k–sgRNA–target DNA map This study EMDB: EMD-24143 Cas12k–sgRNA coordinate This study PDB: 7N3O Cas12k–sgRNA map This study EMDB: EMD-24142 Oligonucleotides DNA primers IDT Table S1 DNA oligos (for structure determination) IDT Table S1 Cas12k sgRNA IDT Table S1 Cas12k sgRNA variants IDT Table S1 Recombinant DNA pDonor_ShCAST_kanR Addgene #127924 pTarget_CAST Addgene #127926 pET28-MKH8SUMO-ShTnsB This paper N/A pET28-MKH8SUMO-ShTnsC This paper N/A pET28-MKH8SUMO-ShTniQ This paper N/A pET-His6-StrepII-TEV LIC-Cas12k This paper N/A pET-His6-StrepII-TEV LIC-Cas12k variant mutants This paper N/A Software and algorithms PyMOL Schrodinger LLC https://pymol.org/2/ UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera/ Phenix Afonine et al., 2018 https://phenix-online.org/ Coot Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ CryoSPARC Punjani et al., 2017 https://cryosparc.com/ (Continued on next page) e2 Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Leginon Suloway et al., 2005 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage CTFFIND4 Rohou and Grigorieff, 2015 https://grigoriefflab.umassmed.edu/ ctffind4 MotionCor2 Zheng et al., 2017 https://github.com/singleparticle/ MotionCor2 PROMALS3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/ promals3d.php ESPript Robert and Gouet, 2014 https://espript.ibcp.fr/ESPript/ESPript/ Sequence Manipulation Suite Stothard, 2000 https://www.bioinformatics.org/sms2/ ident_sim.html GraphPad Prism 8 GraphPad Software https://www.graphpad.com/scientific- software/prism/ National Center for Biotechnology Information (NCBI) Sayers et al., 2021 https://www.ncbi.nlm.nih.gov

    Staining:

    Article Title: Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains BL21-CodonPlus (DE3)-RIL Competent cell Agilent Technologies Cat# 230245 Rosetta (DE3)pLysS Competent cell Novagen Cat# 70956 One Shot PIR1 Chemically Competent cell Thermo fisher scientific Cat# C101010 Stellar Competent Cell Takara Bio Cat# 636763 Chemicals, peptides, and recombinant proteins Terrific Broth (TB) Thermo fisher scientific Cat# BP97285 isopropyl b-D-thiogalactopyranoside (IPTG) Thermo fisher scientific Cat# R0393 Tris-Base Thermo fisher scientific Cat# BP152-1 HEPES Thermo fisher scientific Cat# BP310-1 Sodium chloride Thermo fisher scientific Cat# S271-10 Glycerol Thermo fisher scientific Cat# G33-500 2-Mercaptoethanol Thermo fisher scientific Cat# O3446I-100 Magnesium chloride Thermo fisher scientific Cat# M33-500 PMSF Sigma-Aldrich Cat# P7626 Imidazole Sigma-Aldrich Cat# I202-2KG DTT Thermo fisher scientific Cat# AAJ1539722 EDTA Thermo fisher scientific Cat# AM9261 Potassium Chloride Thermo fisher scientific Cat# P217-500 Magnesium acetate tetrahydrate Millipore Sigma Cat# M5661-250G Agarose Thermo fisher scientific Cat# BP1356-500 SYBR safe DNA Gel Stain Thermo fisher scientific Cat# S33102 Tobacco Etch Virus (TEV) protease Homemade N/A FspI New England Biolabs Cat# R0135L HindIII-HF New England Biolabs Cat# R3104S BseRI New England Biolabs Cat# R0581L BsaI New England Biolabs Cat# R3733S Proteinase K Thermo fisher scientific Cat# EO0491 Benzonase Nuclease Millipore Sigma Cat# E1014-25KU ATP Thermo fisher scientific Cat# BP413-25 Phusion Hot Start II High-Fidelity PCR Master Mix Thermo fisher scientific Cat# F565L Phusion High-Fidelity PCR Master Mix with HF Buffer Thermo fisher scientific Cat# F531L PrimeSTAR Max Premix Takara Bio Cat# R045A In-Fusion HD Cloning Kit Takara Bio Cat# 639650 Q5 Site-Directed Mutagenesis Kit New England Biolabs Cat# E0554 HiScribe T7 Quick High Yield RNA Synthesis Kit New England Biolabs Cat# E2050S QIAprep Spin Miniprep Kit QIAGEN Cat# 27106 MinElute PCR Purification Kit QIAGEN Cat# 28006 ZymoPURE II Plasmid Gigaprep Kit Zymo Research Cat# D4204-B Mini-PROTEAN TGX Precast Gels Bio-rad Cat# 4561086 Amicon Ultra-15 Centrifugal Filter Units 3kDa NMWL Millipore sigma Cat# UFC900324 (Continued on next page) Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Amicon Ultra-15 Centrifugal Filter Units 10kDa NMWL Millipore sigma Cat# UFC901024 Amicon Ultra-15 Centrifugal Filter Units 30kDa NMWL Millipore sigma Cat# UFC903024 Amicon Ultra-15 Centrifugal Filter Units 50kDa NMWL Millipore sigma Cat# UFC905024 NuPAGE LDS Sample Buffer (4X) Thermo fisher scientific Cat# NP0007 Gel Loading Dye, Purple (6X) New England Biolabs Cat# B7024S Bovine Serum Albumin Sigma-Aldrich Cat# A2153-10G Ampicillin, sodium salt Thermo fisher scientific Cat# BP1760-25 Chloramphenicol, 98%, ACROS Organics Thermo fisher scientific Cat# AC227920250 Kanamycin Monosulfate, USP Grade Gold Biotechnology Cat# K-120-5 SnakeSkin Dialysis Tubing, 7K MWCO, 22 mm Thermo fisher scientific Cat# 68700 Diethyl pyrocarbonate Millipore sigma Cat# D5758-25ML Critical commercial assays Ni-NTA agarose resin QIAGEN Cat# 30430 HiTrap Heparin HP affinity column Cytiva Cat# 17040701 RESOURCE Q anion exchange column Cytiva Cat# 17117701 Superdex 200 10/300 column Cytiva Cat# 28990944 InstantBlue Coomassie Protein Stain Abacam Cat# ISB1L Deposited data Cas12k–sgRNA–target DNA coordinate This study PDB: 7N3P Cas12k–sgRNA–target DNA map This study EMDB: EMD-24143 Cas12k–sgRNA coordinate This study PDB: 7N3O Cas12k–sgRNA map This study EMDB: EMD-24142 Oligonucleotides DNA primers IDT Table S1 DNA oligos (for structure determination) IDT Table S1 Cas12k sgRNA IDT Table S1 Cas12k sgRNA variants IDT Table S1 Recombinant DNA pDonor_ShCAST_kanR Addgene #127924 pTarget_CAST Addgene #127926 pET28-MKH8SUMO-ShTnsB This paper N/A pET28-MKH8SUMO-ShTnsC This paper N/A pET28-MKH8SUMO-ShTniQ This paper N/A pET-His6-StrepII-TEV LIC-Cas12k This paper N/A pET-His6-StrepII-TEV LIC-Cas12k variant mutants This paper N/A Software and algorithms PyMOL Schrodinger LLC https://pymol.org/2/ UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera/ Phenix Afonine et al., 2018 https://phenix-online.org/ Coot Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ CryoSPARC Punjani et al., 2017 https://cryosparc.com/ (Continued on next page) e2 Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Leginon Suloway et al., 2005 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage CTFFIND4 Rohou and Grigorieff, 2015 https://grigoriefflab.umassmed.edu/ ctffind4 MotionCor2 Zheng et al., 2017 https://github.com/singleparticle/ MotionCor2 PROMALS3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/ promals3d.php ESPript Robert and Gouet, 2014 https://espript.ibcp.fr/ESPript/ESPript/ Sequence Manipulation Suite Stothard, 2000 https://www.bioinformatics.org/sms2/ ident_sim.html GraphPad Prism 8 GraphPad Software https://www.graphpad.com/scientific- software/prism/ National Center for Biotechnology Information (NCBI) Sayers et al., 2021 https://www.ncbi.nlm.nih.gov

    Recombinant:

    Article Title: Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains BL21-CodonPlus (DE3)-RIL Competent cell Agilent Technologies Cat# 230245 Rosetta (DE3)pLysS Competent cell Novagen Cat# 70956 One Shot PIR1 Chemically Competent cell Thermo fisher scientific Cat# C101010 Stellar Competent Cell Takara Bio Cat# 636763 Chemicals, peptides, and recombinant proteins Terrific Broth (TB) Thermo fisher scientific Cat# BP97285 isopropyl b-D-thiogalactopyranoside (IPTG) Thermo fisher scientific Cat# R0393 Tris-Base Thermo fisher scientific Cat# BP152-1 HEPES Thermo fisher scientific Cat# BP310-1 Sodium chloride Thermo fisher scientific Cat# S271-10 Glycerol Thermo fisher scientific Cat# G33-500 2-Mercaptoethanol Thermo fisher scientific Cat# O3446I-100 Magnesium chloride Thermo fisher scientific Cat# M33-500 PMSF Sigma-Aldrich Cat# P7626 Imidazole Sigma-Aldrich Cat# I202-2KG DTT Thermo fisher scientific Cat# AAJ1539722 EDTA Thermo fisher scientific Cat# AM9261 Potassium Chloride Thermo fisher scientific Cat# P217-500 Magnesium acetate tetrahydrate Millipore Sigma Cat# M5661-250G Agarose Thermo fisher scientific Cat# BP1356-500 SYBR safe DNA Gel Stain Thermo fisher scientific Cat# S33102 Tobacco Etch Virus (TEV) protease Homemade N/A FspI New England Biolabs Cat# R0135L HindIII-HF New England Biolabs Cat# R3104S BseRI New England Biolabs Cat# R0581L BsaI New England Biolabs Cat# R3733S Proteinase K Thermo fisher scientific Cat# EO0491 Benzonase Nuclease Millipore Sigma Cat# E1014-25KU ATP Thermo fisher scientific Cat# BP413-25 Phusion Hot Start II High-Fidelity PCR Master Mix Thermo fisher scientific Cat# F565L Phusion High-Fidelity PCR Master Mix with HF Buffer Thermo fisher scientific Cat# F531L PrimeSTAR Max Premix Takara Bio Cat# R045A In-Fusion HD Cloning Kit Takara Bio Cat# 639650 Q5 Site-Directed Mutagenesis Kit New England Biolabs Cat# E0554 HiScribe T7 Quick High Yield RNA Synthesis Kit New England Biolabs Cat# E2050S QIAprep Spin Miniprep Kit QIAGEN Cat# 27106 MinElute PCR Purification Kit QIAGEN Cat# 28006 ZymoPURE II Plasmid Gigaprep Kit Zymo Research Cat# D4204-B Mini-PROTEAN TGX Precast Gels Bio-rad Cat# 4561086 Amicon Ultra-15 Centrifugal Filter Units 3kDa NMWL Millipore sigma Cat# UFC900324 (Continued on next page) Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Amicon Ultra-15 Centrifugal Filter Units 10kDa NMWL Millipore sigma Cat# UFC901024 Amicon Ultra-15 Centrifugal Filter Units 30kDa NMWL Millipore sigma Cat# UFC903024 Amicon Ultra-15 Centrifugal Filter Units 50kDa NMWL Millipore sigma Cat# UFC905024 NuPAGE LDS Sample Buffer (4X) Thermo fisher scientific Cat# NP0007 Gel Loading Dye, Purple (6X) New England Biolabs Cat# B7024S Bovine Serum Albumin Sigma-Aldrich Cat# A2153-10G Ampicillin, sodium salt Thermo fisher scientific Cat# BP1760-25 Chloramphenicol, 98%, ACROS Organics Thermo fisher scientific Cat# AC227920250 Kanamycin Monosulfate, USP Grade Gold Biotechnology Cat# K-120-5 SnakeSkin Dialysis Tubing, 7K MWCO, 22 mm Thermo fisher scientific Cat# 68700 Diethyl pyrocarbonate Millipore sigma Cat# D5758-25ML Critical commercial assays Ni-NTA agarose resin QIAGEN Cat# 30430 HiTrap Heparin HP affinity column Cytiva Cat# 17040701 RESOURCE Q anion exchange column Cytiva Cat# 17117701 Superdex 200 10/300 column Cytiva Cat# 28990944 InstantBlue Coomassie Protein Stain Abacam Cat# ISB1L Deposited data Cas12k–sgRNA–target DNA coordinate This study PDB: 7N3P Cas12k–sgRNA–target DNA map This study EMDB: EMD-24143 Cas12k–sgRNA coordinate This study PDB: 7N3O Cas12k–sgRNA map This study EMDB: EMD-24142 Oligonucleotides DNA primers IDT Table S1 DNA oligos (for structure determination) IDT Table S1 Cas12k sgRNA IDT Table S1 Cas12k sgRNA variants IDT Table S1 Recombinant DNA pDonor_ShCAST_kanR Addgene #127924 pTarget_CAST Addgene #127926 pET28-MKH8SUMO-ShTnsB This paper N/A pET28-MKH8SUMO-ShTnsC This paper N/A pET28-MKH8SUMO-ShTniQ This paper N/A pET-His6-StrepII-TEV LIC-Cas12k This paper N/A pET-His6-StrepII-TEV LIC-Cas12k variant mutants This paper N/A Software and algorithms PyMOL Schrodinger LLC https://pymol.org/2/ UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera/ Phenix Afonine et al., 2018 https://phenix-online.org/ Coot Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ CryoSPARC Punjani et al., 2017 https://cryosparc.com/ (Continued on next page) e2 Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Leginon Suloway et al., 2005 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage CTFFIND4 Rohou and Grigorieff, 2015 https://grigoriefflab.umassmed.edu/ ctffind4 MotionCor2 Zheng et al., 2017 https://github.com/singleparticle/ MotionCor2 PROMALS3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/ promals3d.php ESPript Robert and Gouet, 2014 https://espript.ibcp.fr/ESPript/ESPript/ Sequence Manipulation Suite Stothard, 2000 https://www.bioinformatics.org/sms2/ ident_sim.html GraphPad Prism 8 GraphPad Software https://www.graphpad.com/scientific- software/prism/ National Center for Biotechnology Information (NCBI) Sayers et al., 2021 https://www.ncbi.nlm.nih.gov

    Variant Assay:

    Article Title: Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains BL21-CodonPlus (DE3)-RIL Competent cell Agilent Technologies Cat# 230245 Rosetta (DE3)pLysS Competent cell Novagen Cat# 70956 One Shot PIR1 Chemically Competent cell Thermo fisher scientific Cat# C101010 Stellar Competent Cell Takara Bio Cat# 636763 Chemicals, peptides, and recombinant proteins Terrific Broth (TB) Thermo fisher scientific Cat# BP97285 isopropyl b-D-thiogalactopyranoside (IPTG) Thermo fisher scientific Cat# R0393 Tris-Base Thermo fisher scientific Cat# BP152-1 HEPES Thermo fisher scientific Cat# BP310-1 Sodium chloride Thermo fisher scientific Cat# S271-10 Glycerol Thermo fisher scientific Cat# G33-500 2-Mercaptoethanol Thermo fisher scientific Cat# O3446I-100 Magnesium chloride Thermo fisher scientific Cat# M33-500 PMSF Sigma-Aldrich Cat# P7626 Imidazole Sigma-Aldrich Cat# I202-2KG DTT Thermo fisher scientific Cat# AAJ1539722 EDTA Thermo fisher scientific Cat# AM9261 Potassium Chloride Thermo fisher scientific Cat# P217-500 Magnesium acetate tetrahydrate Millipore Sigma Cat# M5661-250G Agarose Thermo fisher scientific Cat# BP1356-500 SYBR safe DNA Gel Stain Thermo fisher scientific Cat# S33102 Tobacco Etch Virus (TEV) protease Homemade N/A FspI New England Biolabs Cat# R0135L HindIII-HF New England Biolabs Cat# R3104S BseRI New England Biolabs Cat# R0581L BsaI New England Biolabs Cat# R3733S Proteinase K Thermo fisher scientific Cat# EO0491 Benzonase Nuclease Millipore Sigma Cat# E1014-25KU ATP Thermo fisher scientific Cat# BP413-25 Phusion Hot Start II High-Fidelity PCR Master Mix Thermo fisher scientific Cat# F565L Phusion High-Fidelity PCR Master Mix with HF Buffer Thermo fisher scientific Cat# F531L PrimeSTAR Max Premix Takara Bio Cat# R045A In-Fusion HD Cloning Kit Takara Bio Cat# 639650 Q5 Site-Directed Mutagenesis Kit New England Biolabs Cat# E0554 HiScribe T7 Quick High Yield RNA Synthesis Kit New England Biolabs Cat# E2050S QIAprep Spin Miniprep Kit QIAGEN Cat# 27106 MinElute PCR Purification Kit QIAGEN Cat# 28006 ZymoPURE II Plasmid Gigaprep Kit Zymo Research Cat# D4204-B Mini-PROTEAN TGX Precast Gels Bio-rad Cat# 4561086 Amicon Ultra-15 Centrifugal Filter Units 3kDa NMWL Millipore sigma Cat# UFC900324 (Continued on next page) Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Amicon Ultra-15 Centrifugal Filter Units 10kDa NMWL Millipore sigma Cat# UFC901024 Amicon Ultra-15 Centrifugal Filter Units 30kDa NMWL Millipore sigma Cat# UFC903024 Amicon Ultra-15 Centrifugal Filter Units 50kDa NMWL Millipore sigma Cat# UFC905024 NuPAGE LDS Sample Buffer (4X) Thermo fisher scientific Cat# NP0007 Gel Loading Dye, Purple (6X) New England Biolabs Cat# B7024S Bovine Serum Albumin Sigma-Aldrich Cat# A2153-10G Ampicillin, sodium salt Thermo fisher scientific Cat# BP1760-25 Chloramphenicol, 98%, ACROS Organics Thermo fisher scientific Cat# AC227920250 Kanamycin Monosulfate, USP Grade Gold Biotechnology Cat# K-120-5 SnakeSkin Dialysis Tubing, 7K MWCO, 22 mm Thermo fisher scientific Cat# 68700 Diethyl pyrocarbonate Millipore sigma Cat# D5758-25ML Critical commercial assays Ni-NTA agarose resin QIAGEN Cat# 30430 HiTrap Heparin HP affinity column Cytiva Cat# 17040701 RESOURCE Q anion exchange column Cytiva Cat# 17117701 Superdex 200 10/300 column Cytiva Cat# 28990944 InstantBlue Coomassie Protein Stain Abacam Cat# ISB1L Deposited data Cas12k–sgRNA–target DNA coordinate This study PDB: 7N3P Cas12k–sgRNA–target DNA map This study EMDB: EMD-24143 Cas12k–sgRNA coordinate This study PDB: 7N3O Cas12k–sgRNA map This study EMDB: EMD-24142 Oligonucleotides DNA primers IDT Table S1 DNA oligos (for structure determination) IDT Table S1 Cas12k sgRNA IDT Table S1 Cas12k sgRNA variants IDT Table S1 Recombinant DNA pDonor_ShCAST_kanR Addgene #127924 pTarget_CAST Addgene #127926 pET28-MKH8SUMO-ShTnsB This paper N/A pET28-MKH8SUMO-ShTnsC This paper N/A pET28-MKH8SUMO-ShTniQ This paper N/A pET-His6-StrepII-TEV LIC-Cas12k This paper N/A pET-His6-StrepII-TEV LIC-Cas12k variant mutants This paper N/A Software and algorithms PyMOL Schrodinger LLC https://pymol.org/2/ UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera/ Phenix Afonine et al., 2018 https://phenix-online.org/ Coot Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ CryoSPARC Punjani et al., 2017 https://cryosparc.com/ (Continued on next page) e2 Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Leginon Suloway et al., 2005 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage CTFFIND4 Rohou and Grigorieff, 2015 https://grigoriefflab.umassmed.edu/ ctffind4 MotionCor2 Zheng et al., 2017 https://github.com/singleparticle/ MotionCor2 PROMALS3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/ promals3d.php ESPript Robert and Gouet, 2014 https://espript.ibcp.fr/ESPript/ESPript/ Sequence Manipulation Suite Stothard, 2000 https://www.bioinformatics.org/sms2/ ident_sim.html GraphPad Prism 8 GraphPad Software https://www.graphpad.com/scientific- software/prism/ National Center for Biotechnology Information (NCBI) Sayers et al., 2021 https://www.ncbi.nlm.nih.gov

    Software:

    Article Title: Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains BL21-CodonPlus (DE3)-RIL Competent cell Agilent Technologies Cat# 230245 Rosetta (DE3)pLysS Competent cell Novagen Cat# 70956 One Shot PIR1 Chemically Competent cell Thermo fisher scientific Cat# C101010 Stellar Competent Cell Takara Bio Cat# 636763 Chemicals, peptides, and recombinant proteins Terrific Broth (TB) Thermo fisher scientific Cat# BP97285 isopropyl b-D-thiogalactopyranoside (IPTG) Thermo fisher scientific Cat# R0393 Tris-Base Thermo fisher scientific Cat# BP152-1 HEPES Thermo fisher scientific Cat# BP310-1 Sodium chloride Thermo fisher scientific Cat# S271-10 Glycerol Thermo fisher scientific Cat# G33-500 2-Mercaptoethanol Thermo fisher scientific Cat# O3446I-100 Magnesium chloride Thermo fisher scientific Cat# M33-500 PMSF Sigma-Aldrich Cat# P7626 Imidazole Sigma-Aldrich Cat# I202-2KG DTT Thermo fisher scientific Cat# AAJ1539722 EDTA Thermo fisher scientific Cat# AM9261 Potassium Chloride Thermo fisher scientific Cat# P217-500 Magnesium acetate tetrahydrate Millipore Sigma Cat# M5661-250G Agarose Thermo fisher scientific Cat# BP1356-500 SYBR safe DNA Gel Stain Thermo fisher scientific Cat# S33102 Tobacco Etch Virus (TEV) protease Homemade N/A FspI New England Biolabs Cat# R0135L HindIII-HF New England Biolabs Cat# R3104S BseRI New England Biolabs Cat# R0581L BsaI New England Biolabs Cat# R3733S Proteinase K Thermo fisher scientific Cat# EO0491 Benzonase Nuclease Millipore Sigma Cat# E1014-25KU ATP Thermo fisher scientific Cat# BP413-25 Phusion Hot Start II High-Fidelity PCR Master Mix Thermo fisher scientific Cat# F565L Phusion High-Fidelity PCR Master Mix with HF Buffer Thermo fisher scientific Cat# F531L PrimeSTAR Max Premix Takara Bio Cat# R045A In-Fusion HD Cloning Kit Takara Bio Cat# 639650 Q5 Site-Directed Mutagenesis Kit New England Biolabs Cat# E0554 HiScribe T7 Quick High Yield RNA Synthesis Kit New England Biolabs Cat# E2050S QIAprep Spin Miniprep Kit QIAGEN Cat# 27106 MinElute PCR Purification Kit QIAGEN Cat# 28006 ZymoPURE II Plasmid Gigaprep Kit Zymo Research Cat# D4204-B Mini-PROTEAN TGX Precast Gels Bio-rad Cat# 4561086 Amicon Ultra-15 Centrifugal Filter Units 3kDa NMWL Millipore sigma Cat# UFC900324 (Continued on next page) Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Amicon Ultra-15 Centrifugal Filter Units 10kDa NMWL Millipore sigma Cat# UFC901024 Amicon Ultra-15 Centrifugal Filter Units 30kDa NMWL Millipore sigma Cat# UFC903024 Amicon Ultra-15 Centrifugal Filter Units 50kDa NMWL Millipore sigma Cat# UFC905024 NuPAGE LDS Sample Buffer (4X) Thermo fisher scientific Cat# NP0007 Gel Loading Dye, Purple (6X) New England Biolabs Cat# B7024S Bovine Serum Albumin Sigma-Aldrich Cat# A2153-10G Ampicillin, sodium salt Thermo fisher scientific Cat# BP1760-25 Chloramphenicol, 98%, ACROS Organics Thermo fisher scientific Cat# AC227920250 Kanamycin Monosulfate, USP Grade Gold Biotechnology Cat# K-120-5 SnakeSkin Dialysis Tubing, 7K MWCO, 22 mm Thermo fisher scientific Cat# 68700 Diethyl pyrocarbonate Millipore sigma Cat# D5758-25ML Critical commercial assays Ni-NTA agarose resin QIAGEN Cat# 30430 HiTrap Heparin HP affinity column Cytiva Cat# 17040701 RESOURCE Q anion exchange column Cytiva Cat# 17117701 Superdex 200 10/300 column Cytiva Cat# 28990944 InstantBlue Coomassie Protein Stain Abacam Cat# ISB1L Deposited data Cas12k–sgRNA–target DNA coordinate This study PDB: 7N3P Cas12k–sgRNA–target DNA map This study EMDB: EMD-24143 Cas12k–sgRNA coordinate This study PDB: 7N3O Cas12k–sgRNA map This study EMDB: EMD-24142 Oligonucleotides DNA primers IDT Table S1 DNA oligos (for structure determination) IDT Table S1 Cas12k sgRNA IDT Table S1 Cas12k sgRNA variants IDT Table S1 Recombinant DNA pDonor_ShCAST_kanR Addgene #127924 pTarget_CAST Addgene #127926 pET28-MKH8SUMO-ShTnsB This paper N/A pET28-MKH8SUMO-ShTnsC This paper N/A pET28-MKH8SUMO-ShTniQ This paper N/A pET-His6-StrepII-TEV LIC-Cas12k This paper N/A pET-His6-StrepII-TEV LIC-Cas12k variant mutants This paper N/A Software and algorithms PyMOL Schrodinger LLC https://pymol.org/2/ UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera/ Phenix Afonine et al., 2018 https://phenix-online.org/ Coot Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/ personal/pemsley/coot/ CryoSPARC Punjani et al., 2017 https://cryosparc.com/ (Continued on next page) e2 Molecular Cell 81, 4457–4466.e1–e5, November 4, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Leginon Suloway et al., 2005 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage CTFFIND4 Rohou and Grigorieff, 2015 https://grigoriefflab.umassmed.edu/ ctffind4 MotionCor2 Zheng et al., 2017 https://github.com/singleparticle/ MotionCor2 PROMALS3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/ promals3d.php ESPript Robert and Gouet, 2014 https://espript.ibcp.fr/ESPript/ESPript/ Sequence Manipulation Suite Stothard, 2000 https://www.bioinformatics.org/sms2/ ident_sim.html GraphPad Prism 8 GraphPad Software https://www.graphpad.com/scientific- software/prism/ National Center for Biotechnology Information (NCBI) Sayers et al., 2021 https://www.ncbi.nlm.nih.gov



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