Fast cDNA Synthesis Kit from Bio-Rad

University of Toronto
LMP
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BIo-Rad

Product Name:

iScript™ cDNA Synthesis Kit

Catalog Number:

170-8891

Nowadays most researchers are begging to study the transcriptome by evaluating individual gene expression using qPCR platforms. Importantly, making cDNA using various RT-PCR reagents has been the backbone process in most pipelines. Here, I utilize the iScript cDNA kit from Bio-Rad that is quick and very convenient for converting RNA into cDNA for downstream applications.

Experimental Design and Results Summary

Application

RT-PCR

Starting Material

1 µg DNAse-Free RNA from HCT116 cells

Protocol Overview

-You can convert between ~200 ng–1 µg of DNAse-free RNA into cDNA in a 20 µl total reaction. The kit contains the iScript enzyme and a 5x master mix containing all reagents needed for cDNA synthesis (primer, buffer, dNTPs). The RT-PCR takes ~ 30 mins and the cDNA then can be used for any downstream applications (qPCR, etc.)

Tips

Making a mastermix with the kit's mastermix and the enzyme can speed up the process.

Results Summary

The cDNA was utilized for downstream qPCR reactions. CT values form the qPCR were as expected.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Fast kit for making cDNA

The Bad

Expensive, but works!

The Bottom Line

Great kit that allows you to generate cDNA and run a qPCR within an hour if needed.

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