Real-time Quantitative PCR Using SYBR Green Master Mix

October 08, 2018

BlueskyReddit
Boston University
Center for Regenerative Medicine
Postdoctoral Associate

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Quality of Results

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Company:

BIORAD

Product Name:

iTaq Universal SYBR Green Complex

Catalog Number:

172-5121

The aim of this project was to quantify RSP9 expression levels in our smooth muscle cells. The iTaq Universal SYBR Green Complex master mix was used as it was recommended by colleagues.

Experimental Design and Results Summary

Application

Quantitative real-time PCR (qPCR)

Starting Material

Total RNA extracted from rat smooth muscle cells

Protocol Overview

The master mix was used as per the manufacturer's protocol.

Tips

Run melt-curve post RT-qPCR reaction to detect any non-specific binding

Results Summary

Expression of Rsp9 was detected as expected.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Reactions are easy to set up.

The Bad

Takes a long time to run the R-qPCR

The Bottom Line

The master mix is very sensitive and reliable, however, the RT-qPCR reactions take a long time to complete especially with melt curve included in every run.

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