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RT Master Mix for qPCR (gDNA digester plus)

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MCE RT Master Mix for qPCR (gDNA digester plus) is a convenient, ready-to-use formulation for reverse transcription and can eliminate genomic DNA (gDNA) contaminations in RNA samples. The cDNA product can be directly applied as a template in a standard PCR and real time quantitative PCR (qPCR). The 100 rxns is defined as the base specification. All larger sizes correspond to incremental volumes of this base.

  • Storage :

    -20°C, 2 years

    Avoid repetitive freeze-thaw cycles

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Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Description & Advantages

MCE RT Master Mix is a convenient, ready-to-use formulation for reverse transcription. This kit contains gDNA digester which can eliminate gDNA contaminations in RNA samples. The 2× Super RT Mix contains all the reagents necessary for first-strand cDNA synthesis.The optimized system will provide sensitive and reliable cDNA synthesis. Upon completion of the first-strand cDNA synthesis, the cDNA product can be directly applied as a template in a standard PCR and qPCR. MCE SYBR Green qPCR Master Mix (HY-K0501) is highly recommended for detection of the expression levels of interested genes.

 

Features:

•   Super-fast gDNA removal in 2 minutes.

•   Optimized buffer enhances sensitive and reliable cDNA synthesis.

•   Super-efficient reaction even with low template amounts (total RNA: 5 ng -5 μg in a 20 μL reaction).

•   The kit provides both Oligo dT Primer and Random Primer.

Protocol

1. Thaw RNA templates, gDNA digester, 5× gDNA digester Buffer and the 2× Super RT Mix on ice. Mix solutions gently but thoroughly.

2. Prepare the following reaction mixture in a PCR tube on ice. Mix thoroughly, and incubate at 42°C for 2 minutes.

Components Quantity
5× gDNA digester Buffer 2 μL
gDNA digester 1 μL
Total RNA / mRNA 5 ng-5 μg / 5 ng-500 ng
RNase-Free H2O To 10 μL

3. Add 10 μL of 2× Super RT Mix (gDNA digester inhibitor contained) to the mixture from Step 2 (10 μL). Mix the components well and collect by brief centrifugation. Incubate the mixture in a PCR instrument or water bath in the procedure as follows:

Temperature Time
25°C 5 min
42°C 30-60 min
85°C 2 min

Note:

a. For GC rich or structurally complex RNA templates, increasing the RT incubation temperature up to 50°C may improve the yields of cDNA.

b. Stop the reaction by heating at 85°C for 2 minutes followed by chilling on ice.

4. The newly synthesized first-strand cDNA is ready for immediate downstream applications or for long-term storage at -20°C.

Storage

-20°C, 2 years

Avoid repetitive freeze-thaw cycles

Components

Components HY-K0511-100 rxns HY-K0511-500 rxns
gDNA digester 100 μL 100 μL×5
5× gDNA digester Buffer 200 μL 200 μL×5
2× Super RT Mix 1 mL 1 mL×5
RNase-Free H2O 1 mL×2 1 mL×10

Documentation

MOQ
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100 mg

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