Great SYBR Green for qPCR

Johns Hopkins University
Wilmer
Lab Technologist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Qiagen

Product Name:

Rotor-Gene SYBR Green PCR Kit

Catalog Number:

204076

We isolate RNA from whole mouse retinas and then make cDNA to examine expression of various genes. We use this SYBR Green solution to run qPCR on this cDNA and measure changes in expression between different experimental groups and calculate absolute transcript number.

Experimental Design and Results Summary

Application

Run qPCR to measure cDNA levels

Starting Material

RNA extracted from whole mouse retina

Protocol Overview

A master mix containing 10uL of SYBR Green, 8uL of pure water, 1uL of 10mM primer mix was made. 19uL of mix was added to 1uL of each cDNA sample. Solutions were mixed and placed in a Rotor Gene Q cycler and the specific program was run.

Tips

Always use a control gene for normalization

Results Summary

We are able to successfully and reliable measure gene levels from cDNA using this kit and a fluorescent thermo cycler. This data is reproducible with little variation from run to run.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Simple to use and causes little variability

The Bad

Must be protected from light

The Bottom Line

This is a great Sybr-green product for qpcr.

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