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igScript™ Reverse Transcriptase

$60.00$240.00


    10,000 units (200 u/µl) - $60.00 (Cat.# 3342. )

    50,000 units (200 u/µl) - $240.00 (Cat.# 3344. )

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Description

igScript™ Reverse Transcriptase (igScript RT) is a recombinant Moloney Murine Leukemia Virus (MMLV) reverse transcriptase with reduced RNase H activity. It is highly efficient at producing full-length cDNA from long RNA templates. igScript RT is an RNA-directed DNA polymerase which lacks 3´ → 5´ exonuclease activity and is capable of producing cDNA from as little as 50 pg of total RNA for real-time RT-PCR analysis and other applications.

The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).

reverse-transcriptase-pic1

Additional information

Units

10,000 units (200 u/µl), 50,000 units (200 u/µl)

Product Source

The gene encoding recombinant MMLV Reverse Transcriptase with mutated RNase H domain is expressed in E. coli.

Applications

  • First strand cDNA synthesis for PCR or RT-PCR
  • Gene expression data validation by using RT-PCR product Includes

Benefits

  • Recombinant MMLV reverse transcriptase with greatly reduced RNase H activity.
  • Active at temperatures up to 55°C.
  • Highly efficient at producing full-length cDNA from as little as 50 pg of total RNA.

Product Includes

  1.  igScript™ Reverse Transcriptase
  2. 10x igScript™ RT Reaction Buffer

Storage Temperature
–20 °C

1X MMLV Reverse Transcriptase Reaction Buffer
50 mM Tris-HCl, 75 mM KCl, 3 mM MgCl2, 10 mM DTT, pH 8.3 @ 25°C

Storage Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC

Unit Definition
One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid insoluble material in 10 minutes at 37 °C using poly r(A)/oligo (dT) as a substrate.

Heat Inactivation
65°C for 20 minutes.

First Strand cDNA Synthesis Protocol

  1. In a sterile micro-centrifuge tube, add the following components on ice:
    reverse-transcriptase-pic2
  1. Heat the reaction for 5 minutes at 65 °C. Spin down briefly. Place the tube immediately on ice.
  2. Add the following components:

reverse-transcriptase-pic3

  1. If using random hexamers, incubate the reaction at 25 °C for 10 minutes, then proceed to step 5.
  2. Incubate the reaction at 42 °C for 30-60 minutes.
  3. Inactivate the enzyme by incubating at 65 °C for 20 minutes.
  4. Store products at -20 °C or proceed to next step.

* RNase inhibitor can be purchased from any commercial source

igScript™ Reverse Transcriptase

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