cDNA Synthesis for qPCR Experiments

University of Birmingham
Biosciences
Research Assistant

Overall

Quality of Results

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Company:

Bioline

Product Name:

Tetro cDNA Synthesis kit

Catalog Number:

BIO-65042

Bioline's Tetro cDNA synthesis kit was used to synthesis first strand cDNA using Salmonella enterica total RNA to quantify gene expression. A total of 1 microgram RNA was used and at the end of the reaction, I got an yield of 4 microgram of cDNA in the 20 microliter reaction volume. Gene expression experiments with cDNA went well in qPCR.

Experimental Design and Results Summary

Application

cDNA synthesis for qPCR

Starting Material

Salmonella enterica total RNA

Protocol Overview

A total of one microgram RNA from Salmonella enterica total RNA samples were used for cDNA synthesis using random hexamer primers to prime the cDNA synthesis in a 20 microliter reaction volume. Synthesis steps, temperature cycles are followed according to the kit's instructions. At the end of the reaction, quality and quantify of cDNA was checked by Nanodrop spectrophotometer.

Tips

If you have multiple total RNA samples with different quantities, then normalize input quantities to be used across samples. Try to use same amount of input RNA in the reaction.

Results Summary

One microgram total RNA input was used in the cDNA synthesis reaction and yield was very good with 4 microgram. This was sufficient for my all qPCR experiments. I used 1:10 diluted cDNA as template in the qPCR assays.

Additional Notes

None

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Summary

The Good

Protocol is very simple and easy to perform. Not that much expensive.

The Bad

None

The Bottom Line

This kit performs well with respective synthesis of good quality cDNA for qPCR experiments.

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