igScript™ RT-PCR kit

$160.00$590.00


    50 Reactions - $160.00 (Cat.# 4413 )

    200 Reactions - $590.00 (Cat.# 4415 )

Description

igScript™ RT-PCR Kit combines two powerful mixtures: i). 5x igScript™ master mix and ii) Hot start Taq 2x master mix with standard buffer for 2-steps RT-PCR. The two mixtures require minimal handling during reaction setup and offer consistent and robust RT-PCR reactions.

The first strand cDNA synthesis is achieved by using 5x igScript™ master mix which contains igScript™ Reverse Transcriptase, recombinant RNase inhibitor, dNTPs, an optimized buffer, MgCl2 and protein stabilizers. IgScript™ Reverse Transcriptase is a recombinant MMLV reverse transcriptase with reduced RNase H activity and increased thermostability. The kit also provides two optimized primers and nuclease-free water. An anchored Oligo-dT primer forces the primer to anneal to the beginning of the polyA tail and the random hexamer primer mix provides random and consistent priming sites covering the entire RNA templates including both mRNAs and non-polyadenylated RNAs. The kit is highly efficient at producing full-length cDNA from long RNA templates at temperatures between 42-55ºC.

The amplification step features a high quality Hot start Taq DNA Polymerase which offers higher fidelity and better amplification in a master mix format. RT-PCR product can be generated up to 6 kb.

Applications

  • cDNA synthesis followed by gene expression data validation .

Benefits

  • Robust and active for cDNA synthesis 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. 5x igScript™ master mix
  2. Oligo d(T)23 VN primer (50 µM)
  3. Random hexamer primer mix (60 µM)
  4. Hot start Taq 2x master mix
  5. Nuclease free water

Storage Temperature
–20 °C4413 4415

Additional information

Reactions

50 Reactions, 200 Reactions

(A). First strand cDNA synthesis 

  1. In a sterile micro-centrifuge tube, add the following components on ice:

Picture1

  1. If using random hexamers, incubate the reaction mixture at 25°C for 10 minutes, then proceed to step 3.
  2. Incubate the reaction mixture at temperatures between 42°C to 55ºC for 30-60 minutes.
  3. Inactivate the reaction by incubating at 65°C for 20 minutes.
  4. Proceed to PCR amplification step.

(B). PCR amplification

  1. Prepare a reaction mix according to the following table:

Picture2

  1. Mix the reaction mixture thoroughly.
  2. Program the thermal cycler according to the manufacturer’s instructions.
  3. A typical PCR cycling program is outlined in the following table.
  4. Place the PCR tubes in the thermal cycler and start the cycling program.

Picture3

  1. Analyze 5 μl of PCR products by agarose gel electrophoresis.

 

 

igScript RT-PCR Kit

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