m mulv reverse transcriptase new england biolabs  (New England Biolabs)


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    Name:
    M MuLV Reverse Transcriptase
    Description:
    M MuLV Reverse Transcriptase 50 000 units
    Catalog Number:
    m0253l
    Price:
    288
    Size:
    50 000 units
    Category:
    Reverse Transcriptases
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    New England Biolabs m mulv reverse transcriptase new england biolabs
    M MuLV Reverse Transcriptase
    M MuLV Reverse Transcriptase 50 000 units
    https://www.bioz.com/result/m mulv reverse transcriptase new england biolabs/product/New England Biolabs
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    m mulv reverse transcriptase new england biolabs - by Bioz Stars, 2020-09
    99/100 stars

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    Mutagenesis:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. Two-step RT-PCR reactions were performed using either 5 units 3173 Pol, exonuclease negative mutant or 200 Units MMLV RT (NEB). .. RNA was combined with primers and annealed in water at 70°C for 5 minutes followed by incubation on ice.

    Produced:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. The length distribution of the 3173 Pol cDNAs was visibly shorter than that produced by the MMLV RT, although a subset of the 3173 extension products appeared to be so large that they barely entered the gel. .. Incubation of the DNA primer:RNA template complex with the Taq Pol negative control resulted in a structure-dependent 5′-3′ exonuclease cleavage product that migrated at the dye front .

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. Two-step RT-PCR reactions were performed using either 5 units 3173 Pol, exonuclease negative mutant or 200 Units MMLV RT (NEB). .. RNA was combined with primers and annealed in water at 70°C for 5 minutes followed by incubation on ice.

    other:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: The 3173 Pol and MMLV RT were both able to extend the primer to produce faint, nearly full-length products although the 3173 Pol product was detectably longer than that of MMLV RT.

    Activity Assay:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. As an additional test to compare the RT activity of the 3173 Pol to that of MMLV-RT on a complex RNA substrate, a primer specific to bases 714 to 690 of the negative sense RNA MS2 genome was 5′-fluorophore-labeled. .. The labeled cDNA primer was extended using extracted MS2 RNA as a template ( ).

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. Extension products from a 5′-fluorophore-labeled dT20 primer were resolved by denaturing polyacrylamide gel to further demonstrate RT activity and to assess the relative lengths of the extension products of the 3173 Pol and MMLV RT ( ). ..

    Sequencing:

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme
    Article Snippet: .. Of the two longer target sequences tested, only the 821 bp beta-actin sequence (Lane C3) was reverse transcribed by the 3173 Pol and this synthesis appeared less efficient than that of MMLV RT. ..

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  • 99
    New England Biolabs mmlv rt
    Reverse transcriptase assays. A. Fluorogenic assay. RT activity was measured by detection of RNA:DNA heteroduplex by fluorescence of EvaGreen binding. Oligo dT primed poly A was incubated at 37°C and 65°C in the presence of indicated Pol enzymes in manufacturer recommended buffers and dTTP. Fluorescence measurements were obtained every 6 seconds for 10 minutes. The initial slopes from a plot of RFU vs. time in seconds were determined by linear least square regression from 30 to 150 seconds at 37°C and from 30 to 90 seconds at 65°C. Error bars are standard error of regression slope. B. RT primer extension assay. HEX-labeled dT 20 primed poly A was incubated 10 minutes at 37°C and then 10 minutes at 65°C in the presence of indicated Pol enzymes and dTTP in manufacturer recommended buffers. Primer extension products were resolved by 10% denaturing PAGE and imaged on a Molecular Imager FX (Bio-Rad). Left facing triangle indicates migration of unextended dT20 primer and asterisk indicates bromophenol blue dye front. C. RT MS2-specific primer extension. 5′-labeled primer was annealed to MS2 RNA and incubated 10 minutes at 37°C and then 30 minutes at 65°C in the presence of indicated Pol enzymes with dNTPS (N = A,C,G,T) in manufacturer recommended buffers. Primer extension products were resolved by 5% denaturing PAGE. Lane 1 No <t>RNA+MMLV</t> RT; Lane 2: MS2 RNA No RT; Lane 3 MS2 RNA+MMLV RT, Lane 3 MS2 <t>RNA+3173</t> Pol. Molecular weight in bases indicted. Red Arrow: ∼650 base MMLV extension product. Blue Arrow: ∼715 base PyroScript extension product. Green arrow: Non-templated MMLV reaction product.
    Mmlv Rt, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmlv rt/product/New England Biolabs
    Average 99 stars, based on 35 article reviews
    Price from $9.99 to $1999.99
    mmlv rt - by Bioz Stars, 2020-09
    99/100 stars
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    Reverse transcriptase assays. A. Fluorogenic assay. RT activity was measured by detection of RNA:DNA heteroduplex by fluorescence of EvaGreen binding. Oligo dT primed poly A was incubated at 37°C and 65°C in the presence of indicated Pol enzymes in manufacturer recommended buffers and dTTP. Fluorescence measurements were obtained every 6 seconds for 10 minutes. The initial slopes from a plot of RFU vs. time in seconds were determined by linear least square regression from 30 to 150 seconds at 37°C and from 30 to 90 seconds at 65°C. Error bars are standard error of regression slope. B. RT primer extension assay. HEX-labeled dT 20 primed poly A was incubated 10 minutes at 37°C and then 10 minutes at 65°C in the presence of indicated Pol enzymes and dTTP in manufacturer recommended buffers. Primer extension products were resolved by 10% denaturing PAGE and imaged on a Molecular Imager FX (Bio-Rad). Left facing triangle indicates migration of unextended dT20 primer and asterisk indicates bromophenol blue dye front. C. RT MS2-specific primer extension. 5′-labeled primer was annealed to MS2 RNA and incubated 10 minutes at 37°C and then 30 minutes at 65°C in the presence of indicated Pol enzymes with dNTPS (N = A,C,G,T) in manufacturer recommended buffers. Primer extension products were resolved by 5% denaturing PAGE. Lane 1 No RNA+MMLV RT; Lane 2: MS2 RNA No RT; Lane 3 MS2 RNA+MMLV RT, Lane 3 MS2 RNA+3173 Pol. Molecular weight in bases indicted. Red Arrow: ∼650 base MMLV extension product. Blue Arrow: ∼715 base PyroScript extension product. Green arrow: Non-templated MMLV reaction product.

    Journal: PLoS ONE

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme

    doi: 10.1371/journal.pone.0038371

    Figure Lengend Snippet: Reverse transcriptase assays. A. Fluorogenic assay. RT activity was measured by detection of RNA:DNA heteroduplex by fluorescence of EvaGreen binding. Oligo dT primed poly A was incubated at 37°C and 65°C in the presence of indicated Pol enzymes in manufacturer recommended buffers and dTTP. Fluorescence measurements were obtained every 6 seconds for 10 minutes. The initial slopes from a plot of RFU vs. time in seconds were determined by linear least square regression from 30 to 150 seconds at 37°C and from 30 to 90 seconds at 65°C. Error bars are standard error of regression slope. B. RT primer extension assay. HEX-labeled dT 20 primed poly A was incubated 10 minutes at 37°C and then 10 minutes at 65°C in the presence of indicated Pol enzymes and dTTP in manufacturer recommended buffers. Primer extension products were resolved by 10% denaturing PAGE and imaged on a Molecular Imager FX (Bio-Rad). Left facing triangle indicates migration of unextended dT20 primer and asterisk indicates bromophenol blue dye front. C. RT MS2-specific primer extension. 5′-labeled primer was annealed to MS2 RNA and incubated 10 minutes at 37°C and then 30 minutes at 65°C in the presence of indicated Pol enzymes with dNTPS (N = A,C,G,T) in manufacturer recommended buffers. Primer extension products were resolved by 5% denaturing PAGE. Lane 1 No RNA+MMLV RT; Lane 2: MS2 RNA No RT; Lane 3 MS2 RNA+MMLV RT, Lane 3 MS2 RNA+3173 Pol. Molecular weight in bases indicted. Red Arrow: ∼650 base MMLV extension product. Blue Arrow: ∼715 base PyroScript extension product. Green arrow: Non-templated MMLV reaction product.

    Article Snippet: Two-step RT-PCR reactions were performed using either 5 units 3173 Pol, exonuclease negative mutant or 200 Units MMLV RT (NEB).

    Techniques: Activity Assay, Fluorescence, Binding Assay, Incubation, Primer Extension Assay, Labeling, Polyacrylamide Gel Electrophoresis, Migration, Molecular Weight

    RT-PCR detection of human transcript RNAs. Beta-actin, beta2-microglobulin and cyclophilin target sequences of the indicated sizes were amplified from human liver total RNA using the primers described in Table 1 . Shown are products of two step reactions where either MMLV RT or 3173 Pol were used for first strand cDNA synthesis, as indicated. Taq Pol was used for PCR. Products were resolved on a 1% agarose gel.

    Journal: PLoS ONE

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme

    doi: 10.1371/journal.pone.0038371

    Figure Lengend Snippet: RT-PCR detection of human transcript RNAs. Beta-actin, beta2-microglobulin and cyclophilin target sequences of the indicated sizes were amplified from human liver total RNA using the primers described in Table 1 . Shown are products of two step reactions where either MMLV RT or 3173 Pol were used for first strand cDNA synthesis, as indicated. Taq Pol was used for PCR. Products were resolved on a 1% agarose gel.

    Article Snippet: Two-step RT-PCR reactions were performed using either 5 units 3173 Pol, exonuclease negative mutant or 200 Units MMLV RT (NEB).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis

    Two step RT-PCR comparing 3173 Pol and MMLV RT. A. MS2 viral RNA and B., C. total human liver RNA were reverse transcribed using either 3173 Pol or MMLV RT and then PCR amplified using Taq Polymerase. Target amplicons : A. MS2 RNA phage 77 bp amplicon, 2% gel, B. Human beta-actin 144 bp amplicon, 2% gel, C. Human beta-actin 821 bp amplicon, 1% gel. Lanes : 1 , 11 : oligo dT primer; 2–4 , 12–14 : Gene specific primers; 5 , 15 : random hexamers; 6 , 16 : random nonamers; 7 , 17 : No primer plus RT; 8 : No RT enzyme; 9 : PCR No Target Control; 10 : Molecular Weight Marker (MW), 100 bp (50 bp lowest) for Panels A, B and 1000 bp (300, 500, 700 lowest) for Panel C. Correct PCR product size indicated by black triangle.

    Journal: PLoS ONE

    Article Title: Thermostable DNA Polymerase from a Viral Metagenome Is a Potent RT-PCR Enzyme

    doi: 10.1371/journal.pone.0038371

    Figure Lengend Snippet: Two step RT-PCR comparing 3173 Pol and MMLV RT. A. MS2 viral RNA and B., C. total human liver RNA were reverse transcribed using either 3173 Pol or MMLV RT and then PCR amplified using Taq Polymerase. Target amplicons : A. MS2 RNA phage 77 bp amplicon, 2% gel, B. Human beta-actin 144 bp amplicon, 2% gel, C. Human beta-actin 821 bp amplicon, 1% gel. Lanes : 1 , 11 : oligo dT primer; 2–4 , 12–14 : Gene specific primers; 5 , 15 : random hexamers; 6 , 16 : random nonamers; 7 , 17 : No primer plus RT; 8 : No RT enzyme; 9 : PCR No Target Control; 10 : Molecular Weight Marker (MW), 100 bp (50 bp lowest) for Panels A, B and 1000 bp (300, 500, 700 lowest) for Panel C. Correct PCR product size indicated by black triangle.

    Article Snippet: Two-step RT-PCR reactions were performed using either 5 units 3173 Pol, exonuclease negative mutant or 200 Units MMLV RT (NEB).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Molecular Weight, Marker

    Reverse transcription using AHP dUTP. ( A ) RNA template T5 with primer P3. ( B and C ) Twenty percent denaturing PAGE analysis of reactions using AMV and M-MuLV (RNase H-) reverse transcriptases at 42°C for 15 h.

    Journal: Nucleic Acids Research

    Article Title: Efficient enzymatic synthesis and dual-colour fluorescent labelling of DNA probes using long chain azido-dUTP and BCN dyes

    doi: 10.1093/nar/gkw028

    Figure Lengend Snippet: Reverse transcription using AHP dUTP. ( A ) RNA template T5 with primer P3. ( B and C ) Twenty percent denaturing PAGE analysis of reactions using AMV and M-MuLV (RNase H-) reverse transcriptases at 42°C for 15 h.

    Article Snippet: Klenow large fragment, Therminator™ II, M-MuLV (RNase H− ) reverse transcriptase, AMV reverse transcriptase, RNase inhibitor and λ-exonuclease were purchased from New England Biolabs.

    Techniques: Polyacrylamide Gel Electrophoresis