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NEB/ProtoScript? II Reverse Transcriptase/4,000 units/M0368S
  • NEB/ProtoScript? II Reverse Transcriptase/4,000 units/M0368S

NEB/ProtoScript? II Reverse Transcriptase/4,000 units/M0368S

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貨號(hào): M0368S
品牌: NEB
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    • ProtoScript® II Reverse Transcriptaseis a recombinant M-MuLV reverse transcriptasewith reduced RNase H activity and increasedthermostability. It can be used to synthesize firststrand cDNA at higher temperatures than the wildtype M-MuLV. The enzyme is active up to 48°C,providing higher specificity, higher yield of cDNAand more full-length cDNA product up to 12 kb.
      Protoscript II Reverse Transcriptase performs as well as other RNase H–Reverse TranscriptasesProtoscript II Reverse Transcriptase performs as well as other RNase H– Reverse Transcriptases
      Jurkat total RNA (1 μg) was used in a 20 μl first strand cDNA synthesis. Mixtures of all reaction components, except for reverse transcriptase, were held at different temperatures for 3 min. 200 units SuperScript® II (A) or NEB’s ProtoScript II Reverse Transcriptase(B) was added and incubated at the indicated temperature for 50 minutes, followed by heat inactivation for 5 min at 80°C. 1 μl of cDNA was used in a 25 μl PCR using LongAmpTaqMaster Mix (NEB #M0533) for 35–40 cycles.Ladder L is the 2 Log DNA Ladder (NEB #N0469).
      Robust cDNA synthesis is achieved even with longer templatesRobust cDNA synthesis is achieved even with longer templates
      Jurkat total RNA (1 μg) was used in a 20 μl first strand cDNA synthesis with 200 units of NEB ProtoScript II Reverse Transcriptase. Reactions were incubated at 42°C for 50 minutes, followed by heat inactivation for 5 minutes at 80°C. 1 μl of cDNA was used in a 25 μl PCR using LongAmpTaqMaster Mix (NEB #M0533) for 35–40 cycles. Sizes are indicated above gel.
      Generate high quality cDNA even with very low amounts of starting RNAGenerate high quality cDNA even with very low amounts of starting RNA
      Decreasing amounts of Jurkat total RNA (1 μg – 1 pg) were used in 20 μl first strand cDNA synthesis with 200 units of NEB ProtoScript II Reverse Transcriptase. Reactions were incubated at 42°C for 50 minutes, followed by heat inactivation for 5 minutes at 80°C. 1 μl of cDNA was used in a 25 μl PCR using LongAmpTaqMaster Mix (NEB #M0533) for 40 cycles. The target is a 0.6 kb fragment of GAPDH. Ladder L is the 2-Log DNA Ladder (NEB #N0469).
      ProtoScript II Reverse Transcriptasedisplays superior sensitivityProtoScript II Reverse Transcriptase displays superior sensitivity
      Decreasing amounts of luciferase mRNA (109to 102) molecules were converted into cDNA in the presence of 1 ng Jurkat total RNA using 50 units of NEB ProtoScript II Reverse Transcriptasein a total reaction volume of 20 μl. 1/20 of the cDNA product was amplified using SsoAdvanced™ SYBR® Green Supermix. As few as 5 molecules of luciferase mRNA are detectable.

      Product Source

      The gene encoding a mutant M-MuLV Reverse Transcriptase (RNase H–) is expressed in E.coli and purified to near homogeneity.
      This product is related to the following categories:
      RT-PCR Products,
      Reverse Transcriptases & RT-PCR Products,
      cDNA Synthesis & Reverse Transcriptases Products
      This product can be used in the following applications:
      Helicase-dependent Amplification,
      cDNA Synthesis,
      Reverse Transcription (cDNA Synthesis),
      RT-PCR & cDNA Synthesis,
      RT-qPCR, RT-PCR and cDNA Synthesis,
      PCR
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