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RT-PCR help

reverse transcriptasePCR

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#1 molbio1234

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Posted 14 June 2014 - 08:29 PM

I have isolated RNA from T cells (taken from PBMC). I want to perform qRT-PCR and need to know what are the best housekeeping/endogenous controls to use for this assay? I've read conflicting messages from GAPDH and beta actin and prefer to find one that is good for PBMC/T cell work, or know of a panel of housekeeping genes I can assay. I don't have that much RNA to work with (some samples I only have 1 ug or less), so I need some very stable genes that I can use for the control panel.



#2 Tabaluga

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Posted 15 June 2014 - 01:19 AM

I've heard of this website (but never used it so far): http://www.refgenes.org/rg/

Maybe this could help you ?


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#3 Trof

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Posted 17 June 2014 - 07:12 AM

There are no good universal reference genes.

 

Every reference gene is either suitable or unsuitable for the type of material and treatment you use. Some genes may have other general problems (like actin, it has many pseudogenes). 

The best way to find it it to test several candidates on the .

 

See this topic.

http://www.protocol-...daton-for-qpcr/


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