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Moloney mlv reverse transcriptase

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Moloney MLV reverse transcriptase is an enzyme that catalyzes the conversion of single-stranded RNA into complementary DNA (cDNA). It is commonly used in molecular biology and research applications for the reverse transcription of RNA into cDNA for downstream analysis.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (3 mentions) Viia 7 real time pcr system, by Thermo Fisher Scientific (3 mentions) Rnasin, by Promega (3 mentions) Power sybr green master mix, by Thermo Fisher Scientific (2 mentions) Dnase 1, by Thermo Fisher Scientific (2 mentions) Cfx96 c1000 touch real time pcr system, by Bio-Rad (1 mentions) Iq supermix, by Bio-Rad (1 mentions) Random primer, by Thermo Fisher Scientific (1 mentions) Power syber green master mix, by Thermo Fisher Scientific (1 mentions) Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Reverse transcriptase (531 citations) Moloney murine leukemia virus (M-MLV) reverse transcriptase (57 citations) MLV reverse transcriptase (43 citations) Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (22 citations) Moloney Murine Leukaemia virus (M-MLV) reverse transcriptase (4 citations) Moloney murine leukemia virus (M-MLV) reverse transcriptase kit (2 citations)

5 protocols using moloney mlv reverse transcriptase

1

Quantifying T Cell Gene Expression

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Up to 5 × 106 CD4+ T cells were used for RNA extraction with the InviTrap Spin Kit (Stratec Biomedical) according to the manufacturer’s instructions and up to 500 ng of RNA was retro-transcribed using Moloney MLV reverse transcriptase from Invitrogen according to the manufacturer’s instructions. Gene expression analysis were performed in duplicates using IQ supermix from Biorad in CFX96/C1000 Touch Real-Time PCR system, as described in Lusic et al., 2013. Statistical analysis of qPCR data was performed using Graphpad. Taqman assays used were: for MYC Taqman Hs00153408_m1 FAM/MGB and for GAPDH 4310884E VIC/TAMRA.
Lucic B., Chen H.C., Kuzman M., Zorita E., Wegner J., Minneker V., Wang W., Fronza R., Laufs S., Schmidt M., Stadhouders R., Roukos V., Vlahovicek K., Filion G.J, & Lusic M. (2019). Spatially clustered loci with multiple enhancers are frequent targets of HIV-1 integration. Nature Communications, 10, 4059.
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2

Quantitative Real-Time PCR of Murine Tissues

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For qRT-PCR, frozen tissue was homogenized, and RNA was extracted as previously described (63 (link)). cDNA was synthesized from 1 to 4 μg of total RNA (RNA was isolated from 3 independent mice) using random primers (Invitrogen) and Moloney MLV reverse transcriptase (Invitrogen). PCR was performed in triplicate. Primer sequences available upon request.
Gu Y.F., Cohn S., Christie A., McKenzie T., Wolff N., Do Q.N., Madhuranthakam A.J., Pedrosa I., Wang T., Dey A., Busslinger M., Xie X.J., Hammer R.E., McKay R.M., Kapur P, & Brugarolas J. (2017). Modeling Renal Cell Carcinoma in mice: Bap1 and Pbrm1 Inactivation Drive Tumor Grade. Cancer discovery, 7(8), 900-917.
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3

RNA Extraction and cDNA Synthesis Workflow

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Total RNA was purified from 0.5–2 × 106 cells with TRIzol (Life Technologies). 1 μg RNA was treated with DNase I (Life Technologies) for 15 min at room temperature. DNase I was inactivated by addition of EDTA and heating at 65°C for 10 min. To synthesize cDNA, DNase I-treated RNA was incubated with 125 ng of a 5:1 mixture of oligo-dT primers and random hexamer at 68°C for 10 min. RNA/primers were incubated with Moloney MLV reverse transcriptase (Life Technologies), 10 mM DTT, RNAsin (Promega), and 0.5 mM deoxynucleoside triphosphates (dNTP) at 42°C for 1 h, and then heat inactivated at 98°C for 5 min. Real-time PCR (qRT-PCR) was conducted with Power SYBR Green Master Mix (Applied Biosystems) using ViiA 7 Real-Time PCR system (Applied Biosystems).
Liao R., Zheng Y., Liu X., Zhang Y., Seim G., Tanimura N., Wilson G.M., Hematti P., Coon J.J., Fan J., Xu J., Keles S, & Bresnick E.H. (2020). Discovering How Heme Controls Genome Function Through Heme-omics. Cell reports, 31(13), 107832.
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4

Robust RNA Extraction and qRT-PCR Analysis

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Total RNA was purified with TRIzol (Life Technologies). RNA (1 μg) was DNase treated for 15 min and then heated at 65°C for 10 min with 25 mM EDTA. For cDNA synthesis, DNase I-treated RNA was incubated with 125 ng of a 5:1 mixture of oligo-dT primers and random hexamers at 68°C for 10 min. The RNA and primer mixture was incubated with Moloney MLV reverse transcriptase (Life Technologies), 10 mM DTT, RNAsin (Promega), and 0.5 mM deoxynucleoside triphosphates at 42°C for 1 hour followed by heat inactivation at 98°C for 5 min. qRT-PCR was conducted with Power SYBER Green Master Mix (Applied Biosystems) and a ViiA 7 Real-Time PCR system (Applied Biosystems).
Zwifelhofer N.M., Cai X., Liao R., Mao B., Conn D.J., Mehta C., Keles S., Xia Y, & Bresnick E.H. (2020). GATA factor-regulated solute carrier ensemble reveals a nucleoside transporter-dependent differentiation mechanism. PLoS Genetics, 16(12), e1009286.
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5

RNA Isolation and cDNA Synthesis Protocol

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Total RNA was purified from 0.4–2 × 106 cells with TRIzol (Life Technologies). RNA was treated with DNase I (Life Technologies) for 15 min at room temperature. DNase I was inactivated by EDTA and heating at 65°C for 10 min. DNase I-treated RNA was incubated with 125 ng of a 5:1 mixture of oligo (dT) primers and random hexamer at 68°C for 10 min. RNA/primers were incubated with Moloney MLV reverse transcriptase (Life Technologies), 10 mM DTT (Life Technologies), RNAsin (Promega), and 0.5 mM deoxynucleoside triphosphates at 42°C for 1 h, and then heat inacti vated at 98°C for 5 min. Real-time PCR reactions were conducted with Power SYBR Green Master Mix (Applied Biosystems) using ViiA 7 Real-Time PCR system (Applied Biosystems). Primer sequences are described in Table S3A.
Tanimura N., Liao R., Wilson G.M., Dent M.R., Cao M., Burstyn J.N., Hematti P., Liu X., Zhang Y., Zheng Y., Keles S., Xu J., Coon J.J, & Bresnick E.H. (2018). GATA/Heme Multi-omics Reveals a Trace Metal-Dependent Cellular Differentiation Mechanism. Developmental cell, 46(5), 581-594.e4.
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