maxima rnase h minus reverse transcriptase  (Thermo Fisher)

Article Title: Optogenetic Clustering of Human IRE1 Reveals Differential Regulation of Transcription and mRNA Splice Isoform Abundance by the UPR
Article Snippet: .. The reaction was then quickly mixed with 0.5 μl RNaseOUT (Thermo Fisher #10777019), 1 μl of 20mM strand-switching primer, 1 μl Maxima H Minus Reverse Transcriptase (Thermo Fisher EP0751), and 4 μl Maxima H Minus buffer. ..

Article Title: Supporting Information for mitoSplitter: A mitochondrial variants-based method for efficient demultiplexing of pooled single-cell RNA-seq
Article Snippet: .. In brief, purified mitochondrial RNA (5 μL) was subject to reverse transcription by adding 2 μL 10 μM oligo-dT (ordered from Sangon, SI Appendix, Table S15), 2 μL 10 mM dNTPs (TransGen Biotech, AD101-11), 4 μL 5×RT buffer, 1 μL Maxima H Minus reverse transcriptase (Thermo Scientific, EP0752), 1 μL 50 μM Template Switch oligo (ordered from Sangon, SI Appendix, Table S15), 1 μL RNase Inhibitor (TransGen Biotech, AI101-02), 2 μL 10 mM GTP (Invitrogen, R0461), and 2 μL 50% PEG-8000 (Beyotime, R0056). ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Decoding the Transcriptional Complexity of the Human BRCA2 DNA Repair Gene Using Hybrid-seq.
Article Snippet: Subsequently, mRNA enrichment from 5 μg of each total RNA sample was achieved using the NEBNext® Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, Inc.). .. Reverse transcription (RT) was performed in 20 μl reaction volumes using 100 ng of mRNA from each cell line, oligo-dT20 as the primer, and Maxima H Minus Reverse Transcriptase (InvitrogenTM, Thermo Fisher Scientific Inc.). ..

Article Title: FAM72A promotes UNG2 degradation and mutagenesis in human cancer cells.
Article Snippet: Briefly, total RNA extraction was performed using either Trizol (Invitrogen) or Purelink RNA mini kit (Invitrogen, cat# 12183018 A) according to the manufacture’s protocol. .. Unless otherwise noted, one microgram of RNA has been used to synthesize complementary DNA (cDNA) using Maxima H Minus reverse transcriptase (ThermoFisher) and oligo-dT primers. .. The cDNA samples were synthesized using PowerUp SYBR Green Master Mix (Applied Biosystems) according to the manufactures protocols.

Article Title: Nanoscale 3D DNA tracing in non-denatured cells resolves the Cohesin-dependent loop architecture of the genome in situ
Article Snippet: .. At this point, 1 μl of this sample was set-aside for quality control. ssDNA amplification from template RNA was performed by reverse transcription, using: 150 μl of unpurified in vitro transcription reaction, 1.7 mM dNTP mix, 19 μM forward primer, 240 U RNAsin Plus RNAse inhibitor (Promega), 1200 U Maxima H Minus Reverse Transcriptase (ThermoFisher), 60 μl of 5x Maxima Buffer and water up to 300 μl. ..

Article Title: Supporting Information for Identification of secretory autophagy as a novel mechanism modulating activity- induced synaptic remodeling
Article Snippet: To determine GFP-Atg8a, mCherry-Atg8a, and GFP-Lamp1 intensities, type Ib boutons were circled based on HRP signal, and the average intensities (normalized to the bouton area) were measured using Image J. Quantitative RT-PCR Total RNA was extracted from 10 larval brains using TRIzol reagent (Invitrogen). .. Subsequently, 2 μg of total RNA was used for cDNA synthesis using the Maxima H Minus Reverse Transcriptase (Thermo Scientific). .. Quantitative RT-PCR using SYBR Green reagent (Applied Biosystems) was carried out to detect mRNA sequences for either snap29, rab8, or sec22.

Article Title: Decoding the Transcriptional Complexity of the Human BRCA2 DNA Repair Gene Using Hybrid-seq.
Article Snippet: .. Subsequently, the cDNA synthesis mixture was completed by adding 4 μl of 5X RT Buffer, 1 μl of dNTP mix (10 mM each), 0.5 μl of RNaseOUT inhibitor (20 U; InvitrogenTM, Thermo Fisher Scientific Inc.), and 1 μl of Maxima H Minus Reverse Transcriptase (200 U; InvitrogenTM, Thermo Fisher Scientific Inc.). ..

Article Title: Supporting Information for mitoSplitter: A mitochondrial variants-based method for efficient demultiplexing of pooled single-cell RNA-seq
Article Snippet: .. In brief, purified mitochondrial RNA (5 μL) was subject to reverse transcription by adding 2 μL 10 μM oligo-dT (ordered from Sangon, SI Appendix, Table S15), 2 μL 10 mM dNTPs (TransGen Biotech, AD101-11), 4 μL 5×RT buffer, 1 μL Maxima H Minus reverse transcriptase (Thermo Scientific, EP0752), 1 μL 50 μM Template Switch oligo (ordered from Sangon, SI Appendix, Table S15), 1 μL RNase Inhibitor (TransGen Biotech, AI101-02), 2 μL 10 mM GTP (Invitrogen, R0461), and 2 μL 50% PEG-8000 (Beyotime, R0056). ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout
Article Snippet: RNA was extracted from samples using TRIzol (Invitrogen 15596018) and an RNA extraction kit following the manufacturer’s recommended protocol (Zymo R2062). .. RNA concentration and purity were measured on a spectrophotometer. cDNA was produced from 1 μg of RNA per sample, using Maxima H Minus Reverse Transcriptase (Thermo Scientific EP0752) and following the manufacturer’s recommendations. qPCR reactions were prepared in a 384-well plate (Applied Biosystems 4309849) with 1:10 dilution of cDNA, 2x PowerUp SYBR Green Master Mix (Applied Biosystems A25741) and 1 μM each of forward and reverse primers. qPCRs were run on QuantStudio 7 (Applied Biosystems 4485701) and analyzed using the ΔΔCt method, normalized to ACTB and S18 rRNA. ..

Article Title: Nanoscale 3D DNA tracing in non-denatured cells resolves the Cohesin-dependent loop architecture of the genome in situ
Article Snippet: .. At this point, 1 μl of this sample was set-aside for quality control. ssDNA amplification from template RNA was performed by reverse transcription, using: 150 μl of unpurified in vitro transcription reaction, 1.7 mM dNTP mix, 19 μM forward primer, 240 U RNAsin Plus RNAse inhibitor (Promega), 1200 U Maxima H Minus Reverse Transcriptase (ThermoFisher), 60 μl of 5x Maxima Buffer and water up to 300 μl. ..

Article Title: Nanoscale 3D DNA tracing in non-denatured cells resolves the Cohesin-dependent loop architecture of the genome in situ
Article Snippet: .. At this point, 1 μl of this sample was set-aside for quality control. ssDNA amplification from template RNA was performed by reverse transcription, using: 150 μl of unpurified in vitro transcription reaction, 1.7 mM dNTP mix, 19 μM forward primer, 240 U RNAsin Plus RNAse inhibitor (Promega), 1200 U Maxima H Minus Reverse Transcriptase (ThermoFisher), 60 μl of 5x Maxima Buffer and water up to 300 μl. ..

Article Title: Nanoscale 3D DNA tracing in non-denatured cells resolves the Cohesin-dependent loop architecture of the genome in situ
Article Snippet: .. At this point, 1 μl of this sample was set-aside for quality control. ssDNA amplification from template RNA was performed by reverse transcription, using: 150 μl of unpurified in vitro transcription reaction, 1.7 mM dNTP mix, 19 μM forward primer, 240 U RNAsin Plus RNAse inhibitor (Promega), 1200 U Maxima H Minus Reverse Transcriptase (ThermoFisher), 60 μl of 5x Maxima Buffer and water up to 300 μl. ..

Article Title: Supporting Information for Identification of secretory autophagy as a novel mechanism modulating activity- induced synaptic remodeling
Article Snippet: To determine GFP-Atg8a, mCherry-Atg8a, and GFP-Lamp1 intensities, type Ib boutons were circled based on HRP signal, and the average intensities (normalized to the bouton area) were measured using Image J. Quantitative RT-PCR Total RNA was extracted from 10 larval brains using TRIzol reagent (Invitrogen). .. Subsequently, 2 μg of total RNA was used for cDNA synthesis using the Maxima H Minus Reverse Transcriptase (Thermo Scientific). .. Quantitative RT-PCR using SYBR Green reagent (Applied Biosystems) was carried out to detect mRNA sequences for either snap29, rab8, or sec22.

Article Title: Decoding the Transcriptional Complexity of the Human BRCA2 DNA Repair Gene Using Hybrid-seq.
Article Snippet: .. Subsequently, the cDNA synthesis mixture was completed by adding 4 μl of 5X RT Buffer, 1 μl of dNTP mix (10 mM each), 0.5 μl of RNaseOUT inhibitor (20 U; InvitrogenTM, Thermo Fisher Scientific Inc.), and 1 μl of Maxima H Minus Reverse Transcriptase (200 U; InvitrogenTM, Thermo Fisher Scientific Inc.). ..