Real-Time PCR Reaction for Glioma Stem Cell cDNA

Ludwig Cancer Research
Centre for Cellular and Molecular Medicine
Postdoctoral Researcher

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Company:

Bio-Rad

Product Name:

iTaq™ Universal SYBR® Green Supermix

Catalog Number:

1725124

Our aim is to examine the expression of genes involved in regulation of tumor growth. RNA was isolated from approx 3 million cells using RNAeasy extraction kit from qiagen. 3ug of RNA was used to prepare cDNA using RNA to cDNA ecodry premix from Takara biosciences. cDNA was diluted with molecular biology grade water to obtain a concentration of 10 ng/ul. Real time PCR reaction (20uL) was set up by adding 10 ul sybr green mix, 8 ul of water, 0.5ul forwar primer (10uM), 0.5 uL reverse primer (10uM), 1uL of cDNA (diluted). All the samples for ran in triplicates.

Experimental Design and Results Summary

Application

Real time PCR

Starting Material

Glioma stem cells cDNA

Tips

N/A

Results Summary

Observed detection of few genes which are expressed in low levels in glioma stem cell lines

DOI or PMID #

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Summary

The Good

Efficient detection of least expressing genes

The Bad

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The Bottom Line

Excellent product for real time PCR. Ready to use mix will make the PCR reaction easy to setup and error between triplicates will be reduced significantly

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