5x first strand buffer  (Thermo Fisher)


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    Structured Review

    Thermo Fisher 5x first strand buffer
    5x First Strand Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5x first strand buffer/product/Thermo Fisher
    Average 92 stars, based on 138 article reviews
    Price from $9.99 to $1999.99
    5x first strand buffer - by Bioz Stars, 2020-09
    92/100 stars

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    Related Articles

    High Performance Liquid Chromatography:

    Article Title: Strand-specific RNA-seq applied to malaria samples
    Article Snippet: .. Parasite cultures grown in complete RPMI medium at 5 % hematocrit ( see ) TRIzol® LS Reagent (Invitrogen™) pre-warmed at 37°C Chloroform Isopropanol pre-chilled on ice Nuclease-free non-DEPC water DNAse I RNAse-free (Ambion®) Deionized formamide Formaldehyde 37 % 10X MOPS EDTA buffer pH 7.0 Glycerol 50 % Bromophenol blue powder Ethidium bromide 20 mg/mL GenElute™ mRNA Miniprep Kit (Sigma-Aldrich) 5X RNA storage solution (Ambion) HPLC-purified random hexamers and Anchored OligodT(20) SuperScript® VILO™ cDNA synthesis kit (Invitrogen™) DNA Clean & Concentrator™ (Zymo Research) 5X First Strand Buffer (Invitrogen™): 250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 5X Second Strand Buffer (Invitrogen™): 100 mM Tris-HCl (pH 6.9), 450 mM KCl, 23 mM MgCl2 , 0.75 mM β-NAD+ , 50 mM (NH4 )2 SO4 Set of dATP, dGTP, dCTP, and dUTP E. coli DNA Polymerase I 10 U/μL (Invitrogen™) E. coli DNA Ligase 10 U/μL (Invitrogen™) E. coli DNA RNase H 2 U/μL (Invitrogen™) 0.1 M DTT dsDNA Shearase™ (Zymo Research) Encore™ NGS Library System I (NuGEN®) Same-day 70 % ethanol in nuclease-free water USER™ Enzyme (New England Biolabs®) 1X TE buffer pH 8.0 ..

    Centrifugation:

    Article Title: SNP variation in the promoter of the PRKAG3 gene and association with meat quality traits in pig
    Article Snippet: .. The contents were collected by a brief centrifugation before adding 4 μl 5X first strand buffer, 2 μl 0.1 M DTT, 1 μl SUPERase-In (Ambion, Foster City) and 1 μl of Superscript III RNase H reverse transcriptase (200 u/μl) (Invitrogen, Carlsbad, CA). ..

    In Vitro:

    Article Title: Optimized amplification and single-cell analysis identify GnRH-mediated activation of Rap1b in primary rat gonadotropes
    Article Snippet: .. Briefly, 100 ng total RNA was incubated in a 10-μl volume containing 2 μl T7-Oligo(dT) primer, 2 μl 5X first strand buffer, 1 μl 0.1 M DTT, 0.5 μl RNase inhibitor, 0.5 μl 10 mM dNTP, and 1 μl SuperScript II (Invitrogen) at 42 ºC for 1 h. Second strand cDNA synthesis was carried out by incubating 10 μl first strand cDNA products with 4.8 μl water, 4 μl 17.5 mM MgCl2 , 0.4 μl 10 mM dNTP, 0.6 μl E. coli DNA Polymerase I, and 0.2 μl RNase H at 16ºC for 2 h. In vitro transcription was performed using the MegaScript T7 Kit (Ambion). .. Second strand cDNA products (20 μl) were incubated with 5 μl 10X Reaction buffer, 5 μl each of ATP, CTP, UTP and GTP solutions, and 5 μl Enzyme Mix at 37ºC for 16 h. The cRNA generated was purified using an RNeasy kit (Qiagen) and the manufacturer’s RNA cleanup protocol. cRNA was subsequently subjected to a second cycle of T7 amplification.

    Next-Generation Sequencing:

    Article Title: Strand-specific RNA-seq applied to malaria samples
    Article Snippet: .. Parasite cultures grown in complete RPMI medium at 5 % hematocrit ( see ) TRIzol® LS Reagent (Invitrogen™) pre-warmed at 37°C Chloroform Isopropanol pre-chilled on ice Nuclease-free non-DEPC water DNAse I RNAse-free (Ambion®) Deionized formamide Formaldehyde 37 % 10X MOPS EDTA buffer pH 7.0 Glycerol 50 % Bromophenol blue powder Ethidium bromide 20 mg/mL GenElute™ mRNA Miniprep Kit (Sigma-Aldrich) 5X RNA storage solution (Ambion) HPLC-purified random hexamers and Anchored OligodT(20) SuperScript® VILO™ cDNA synthesis kit (Invitrogen™) DNA Clean & Concentrator™ (Zymo Research) 5X First Strand Buffer (Invitrogen™): 250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 5X Second Strand Buffer (Invitrogen™): 100 mM Tris-HCl (pH 6.9), 450 mM KCl, 23 mM MgCl2 , 0.75 mM β-NAD+ , 50 mM (NH4 )2 SO4 Set of dATP, dGTP, dCTP, and dUTP E. coli DNA Polymerase I 10 U/μL (Invitrogen™) E. coli DNA Ligase 10 U/μL (Invitrogen™) E. coli DNA RNase H 2 U/μL (Invitrogen™) 0.1 M DTT dsDNA Shearase™ (Zymo Research) Encore™ NGS Library System I (NuGEN®) Same-day 70 % ethanol in nuclease-free water USER™ Enzyme (New England Biolabs®) 1X TE buffer pH 8.0 ..

    Incubation:

    Article Title: Pair-wise comparison analysis of differential expression of mRNAs in early and advanced stage primary colorectal adenocarcinomas
    Article Snippet: .. The reaction mixture was then incubated at 80°C for 3 min, followed by chilling on ice for another 2 min. Next, 4 µL of 5X first-strand buffer (Invitrogen), 5 µL of 2 mM dNTP (Fermentas), 0.5 µL of 40 U/µL RNaseOUT recombinant RNase inhibitor (Invitrogen) and 1 µL of 200 U/µL M-MLV reverse transcriptase (Invitrogen) were added to the mixture. ..

    Article Title: Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)
    Article Snippet: .. The mixture was incubated at 65°C for 5 min and kept on ice for 3 min. A total of 6 μ L of master mix composed of 4.5 μ L 5X First-Strand Buffer, 1 μ L 0.1 M DTT, 0.25 μ L (50 U) of SuperScript III RT (Invitrogen Corp., CA), and 0.25 μ L of RNase inhibitor (10 U, Promega, WI) was added. .. The reaction was performed in an Eppendorf Gradient cycler using the program: 25°C for 5 min, 50°C for 60 min and 70°C for 15 min. cDNA was then diluted 1 : 4 (v : v) with DNase-/RNase-free water.

    Article Title: Optimized amplification and single-cell analysis identify GnRH-mediated activation of Rap1b in primary rat gonadotropes
    Article Snippet: .. Briefly, 100 ng total RNA was incubated in a 10-μl volume containing 2 μl T7-Oligo(dT) primer, 2 μl 5X first strand buffer, 1 μl 0.1 M DTT, 0.5 μl RNase inhibitor, 0.5 μl 10 mM dNTP, and 1 μl SuperScript II (Invitrogen) at 42 ºC for 1 h. Second strand cDNA synthesis was carried out by incubating 10 μl first strand cDNA products with 4.8 μl water, 4 μl 17.5 mM MgCl2 , 0.4 μl 10 mM dNTP, 0.6 μl E. coli DNA Polymerase I, and 0.2 μl RNase H at 16ºC for 2 h. In vitro transcription was performed using the MegaScript T7 Kit (Ambion). .. Second strand cDNA products (20 μl) were incubated with 5 μl 10X Reaction buffer, 5 μl each of ATP, CTP, UTP and GTP solutions, and 5 μl Enzyme Mix at 37ºC for 16 h. The cRNA generated was purified using an RNeasy kit (Qiagen) and the manufacturer’s RNA cleanup protocol. cRNA was subsequently subjected to a second cycle of T7 amplification.

    Spectrophotometry:

    Article Title: Protective Effect of Heme Oxygenase-1 on High Glucose-Induced Pancreatic ?-Cell Injury
    Article Snippet: .. After quantification by spectrophotometer (UV/VIS Spectrophotometer ND-1000; Nanodrop, Silmington, DE, USA), 2 µg of RNA was subjected to 85℃ for 3 minutes prior to the addition of 5X First-Strand buffer (Invitrogen), 0.1 M dTT (Invitrogen), 50 µg/µL Random hexamer (Invitrogen), 10 mM dNTP (Invitrogen), RNaseOUT™ (40 U/µL, Invitorogen), and SuperScrip™ II Reverse Transcriptase (200 U/µL, Invitrogen). ..

    Random Hexamer Labeling:

    Article Title: Protective Effect of Heme Oxygenase-1 on High Glucose-Induced Pancreatic ?-Cell Injury
    Article Snippet: .. After quantification by spectrophotometer (UV/VIS Spectrophotometer ND-1000; Nanodrop, Silmington, DE, USA), 2 µg of RNA was subjected to 85℃ for 3 minutes prior to the addition of 5X First-Strand buffer (Invitrogen), 0.1 M dTT (Invitrogen), 50 µg/µL Random hexamer (Invitrogen), 10 mM dNTP (Invitrogen), RNaseOUT™ (40 U/µL, Invitorogen), and SuperScrip™ II Reverse Transcriptase (200 U/µL, Invitrogen). ..

    Recombinant:

    Article Title: Pair-wise comparison analysis of differential expression of mRNAs in early and advanced stage primary colorectal adenocarcinomas
    Article Snippet: .. The reaction mixture was then incubated at 80°C for 3 min, followed by chilling on ice for another 2 min. Next, 4 µL of 5X first-strand buffer (Invitrogen), 5 µL of 2 mM dNTP (Fermentas), 0.5 µL of 40 U/µL RNaseOUT recombinant RNase inhibitor (Invitrogen) and 1 µL of 200 U/µL M-MLV reverse transcriptase (Invitrogen) were added to the mixture. ..

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    Thermo Fisher superscript iii reverse transcriptase
    Microcapillary electrophoresis of total <t>RNA</t> from whole cells and projections. Microcapillary electrophoresis of total RNA (Bioanalyzer RNA pico assay) from whole cells and projections of <t>three</t> biological replicates. Only RNAs > ~150 nt were sequenced for this study. In projections, the band at ~100 nt was striking. By sequencing RNAs within 20–150 nt size range, we found that they were mostly full length and fragmented tRNAs (unpublished data).
    Superscript Iii Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7838 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher first strand buffer
    Microcapillary electrophoresis of total <t>RNA</t> from whole cells and projections. Microcapillary electrophoresis of total RNA (Bioanalyzer RNA pico assay) from whole cells and projections of <t>three</t> biological replicates. Only RNAs > ~150 nt were sequenced for this study. In projections, the band at ~100 nt was striking. By sequencing RNAs within 20–150 nt size range, we found that they were mostly full length and fragmented tRNAs (unpublished data).
    First Strand Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/first strand buffer/product/Thermo Fisher
    Average 94 stars, based on 1151 article reviews
    Price from $9.99 to $1999.99
    first strand buffer - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

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    Microcapillary electrophoresis of total RNA from whole cells and projections. Microcapillary electrophoresis of total RNA (Bioanalyzer RNA pico assay) from whole cells and projections of three biological replicates. Only RNAs > ~150 nt were sequenced for this study. In projections, the band at ~100 nt was striking. By sequencing RNAs within 20–150 nt size range, we found that they were mostly full length and fragmented tRNAs (unpublished data).

    Journal: eLife

    Article Title: Free circular introns with an unusual branchpoint in neuronal projections

    doi: 10.7554/eLife.47809

    Figure Lengend Snippet: Microcapillary electrophoresis of total RNA from whole cells and projections. Microcapillary electrophoresis of total RNA (Bioanalyzer RNA pico assay) from whole cells and projections of three biological replicates. Only RNAs > ~150 nt were sequenced for this study. In projections, the band at ~100 nt was striking. By sequencing RNAs within 20–150 nt size range, we found that they were mostly full length and fragmented tRNAs (unpublished data).

    Article Snippet: RNA was hydrolyzed using 5X first strand buffer (provided with Superscript III reverse transcriptase, ThermoFisher #18080044) at 94°C for 4 min and 50 s and immediately moved to ice.

    Techniques: Electrophoresis, Sequencing