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QuantiSure First Strand cDNA Synthesis Kit

Cat. No.





25 x 20-µl reactions



50 x 20-µl reactions



200 x 20-µl reactions



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QuantiSure First Strand cDNA Synthesis Kit, designed for use in two-step RT-qPCR, provides a fast and efficient procedure for reverse transcription and genomic DNA elimination in your RNA samples. To obtain accurate results in real-time RT-PCR, it is important that only cDNA is amplified and detected. Interference by genomic DNA can be avoided by designing primers or probes that span an exon/exon boundary. However, in cases where this is not possible (e.g., the cDNA is from a single-exon gene, or there exist processed pseudogenes in genomic DNA sequences), it is essential that the starting RNA sample is free of genomic DNA. QuantiSure First Strand cDNA Synthesis Kit combines genomic DNA elimination of RNA samples and subsequent first strand cDNA synthesis in one easy workflow to ensure accuracy of real-time RT-PCR results. Since there is no need to perform a separate DNase digestion and re-purify your RNA samples, your time and costs are saved.




gDNA Removing Buffer (4X)

Buffer for effective elimination of genomic DNA contamination in RNA samples

RT Buffer (5X)

Buffer optimized for reverse transcription; Contains MgCl2, dNTPs, and stabilizers

Enzyme Mix

Mixture of reverse transcriptase, and RNase inhibitor.

Primer Mix

Optimized blend of oligo-dT, and random primers.

RNase-free H2O

Ultrapure quality



The kit is stable for one year when stored in a constant temperature freezer at -20ºC. After thawing, mix the components thoroughly before using. Frequent freezing and thawing is not recommended.  

Quick-Reference Protocol

1. Thaw gDNA Removing Buffer (4X), RT Buffer (5X), Primer Mix, and RNase-free water at room temperature (15–25°C).  Mix each solution by flicking the tubes. Centrifuge briefly, and then store on ice.

2. Prepare a Genomic DNA Elimination Reaction on ice. Mix and then store on ice.   


Volume /Reaction

gDNA Removing Buffer (4 X)

3 µl

Template RNA

Variable (up to 1 µg total RNA)

RNase-free H2O


Total Reaction Volume

12 µl


3. Incubate for 5 min at 37ºC. Then place on ice.

4. Prepare the Reverse Transcription Master Mix on ice.


Volume /Reaction

RT Buffer (5 X)

4 µl

Primer Mix

3 µl

Enzyme Mix

1 µl

Total Volume

8 µl


5. Add 8 µl Reverse Transcription Master Mix to each of Genomic DNA Elimination Reactions from step 3 (12 µl) for a total reaction volume of 20 µl. Mix and then store on ice.

6. Incubate for 15 min at 42ºC.

7. Incubate for 3 min at 95°C to stop the reaction.

8. Place finished reverse-transcription reaction on ice until ready to use in real-time PCR. For long-term storage, place reverse-transcription reaction at -20ºC.