First-Strand cDNA Synthesis Using iScript™ cDNA Synthesis Kit

Uka Tarsadia University

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Quality of Results

Ease-of-Optimization

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Company:

Biorad

Product Name:

iScript™ cDNA Synthesis Kit

Catalog Number:

1708891

The aim of the experiment was to synthesize cDNA from 1 μg RNA isolated from mammalian cells for gene expression study by qPCR using gene specific primers.

Experimental Design and Results Summary

Application

Real-time qPCR

Starting Material

Total RNA

Tips

Total 1 µg–1 pg input RNA is needed

Results Summary

We easily and efficiently synthesize first-strand cDNA by using iScript™ cDNA Synthesis Kit (Biorad, Hercules, CA) and used it for gene expression analysis using real-time qPCR. The kit is sensitive and cDNA yield is very good. The protocol is very easy and less time consuming.

DOI or PMID #

DOI: 10.1111/imm.13538

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Summary

The Good

The kit provides complete cDNA synthesis. The protocol is less time consuming (26 minute protocol)

The Bad

The Kit is bit expensive. Good quality RNA with optimum concentration is needed

The Bottom Line

iScript™ cDNA Synthesis Kit (Biorad, Hercules, CA) is very apt to synthesize first-strand cDNA from total RNA. Moreover, the protocol is very simple and results are consistent. The kit is very useful for detecting low-level target genes. However, good quality RNA with optimum concentration is needed and little optimization may be needed due to different cell types of the experiments.

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