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RT Master Mix for qPCR II (gDNA digester plus)

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MCE RT Master Mix for qPCR II is a convenient, ready-to-use formulation for reverse transcription and can eliminate genomic DNA (gDNA) contaminations in RNA samples. With updated reverse transcriptase, this kit can synthesize cDNA more rapidly, more specifically. 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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To place orders, for customer services and technical support, please contact: MedChemExpress USA

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 5x gDNA digester Mix which can eliminate gDNA contaminations in RNA samples. 4× Super RT Mix contains all the reagents necessary for first-strand cDNA synthesis. In addition, the increased heat resistance of the advanced reverse transcriptase allows reverse transcription to be performed at a higher temperature, increasing the efficiency and specificity of 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:

1. Synthesize cDNA rapidly and specifically.

2. Super-fast gDNA removal in 2 minutes.

3. Optimized buffer enhances sensitive and reliable cDNA synthesis.

4. Super-efficient reaction even with low template amounts (total RNA: 50 pg -5 μg in a 20 μL reaction).

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

Protocol

1. Thaw RNA templates, gDNA digester, 5× gDNA digester Buffer and the 4× 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 3 μL
Total RNA / mRNA 50 pg-5 μg / 50 pg-500 ng
RNase-Free H2O To 15 μL

3. Add 5 μL of 4× Super RT Mix (gDNA digester inhibitor contained) to the mixture from Step 2 (15 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
55°C 15 min
85°C 2 min

Note:

a. For GC rich or structurally complex RNA templates, increasing the RT incubation temperature up to 60°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

Contents HY-K0511A-100 rxns HY-K0511A-500 rxns
5× gDNA digester Mix 300 μL 300 μL × 5
4× Super RT Mix 500 μL 500 μL × 5
RNase-Free H2O 1 mL × 2 1 mL × 10

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MOQ
Minimum order quantity
100 mg

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