SYBR Green for Quantitative Real-Time PCR

March 06, 2020

BlueskyReddit
Centre for Cellular & Molecular Biology
Project JRF
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Applied Biosystems

Product Name:

Power SYBR green PCR matermix

Catalog Number:

4367659

This SYBR green mastermix is ready to use for your quantitative realtime PCR experiments. A minimum of 5uL of master mix is enough for a 10uL reaction volume containing primers and template DNA. Doesn't require much optimization if you are following the recommended primer (50nM to 900nM) and template concentrations (1-100ng). Applied biosystem qPCR machines are highly recommended for the quantification with this SYBR green. However, even Roche machine also did the job in my experiment. My experiment reaction mixture is as follows:SYBR green-5uLForward and reverse primers - 0.2uL each from 100uM stockTemplate cDNA - 1uL (synthesized from 100ng RNA and diluted twice)Molecular biology grade water - 3.6uL

Experimental Design and Results Summary

Application

Absolute and quantitative realtime PCR

Starting Material

cDNA prepared from RNA or genomic DNA

Tips

Take the required amount for running your experiment from the stock to avoid exposing the SYBR to sunlight.

Results Summary

For quantitative PCR, amplification cure and the melting curve need to be referred along with the housekeeping genes and the non-template controls.

DOI or PMID #

N/A

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Summary

The Good

Less optimization required and results are very much reproducible.

The Bad

Nothing, however there are some colorful mastermix available in the market for easy loading in the wells.

The Bottom Line

If someone wants biological answers and doesn't want to spend much time in optimization. Please go for it.

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