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random primers or oligodT


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4 replies to this topic

#1 SF_HK

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Posted 20 March 2013 - 01:05 AM

I'm confused as to use random or oligodT primers. I have to check expression of various egenes in around 130 tissues. Most reasonable kits (with 100+ reactions), use randome hexamer primers.

I checked my 12 genes of interest on cancer cell lines whose RNA was extracted using olidodT primers. I would ideally like to use oligodT primers for my RT-PCR in the clinical tissues, but the kits are very expensive. The cheaper kits use random primers.

Any advice?

#2 Neanderthal

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Posted 20 March 2013 - 03:39 AM

When you checked your 12 genes of interest on cancer cell lines (whose RNA was extracted using olidodT primers) did you come across any problems? Is that the reason you want to use oligodT? If you have stumbled upon any problems, then why can't you sgo ahead with random hexamer primers? I am curious to know if there is any specific reason.

Kart

#3 SF_HK

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Posted 20 March 2013 - 07:07 AM

Thanks. I did not come across any problems. I was asking regarding consistency. I checked the expression of the 12 genes in cell lines whose RNA was extracted using oligodt. Thus I should also use oligodt RNA extraction from my clinical tissues? It would mean inconsistency if the RNA from cell lines is made using oligodt and the RNA from clinical samples using random hexamers?

#4 memari

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Posted 20 March 2013 - 09:21 AM

This one that we use has both of them (olidodT + random hexamers) as a Mix in its buffer:

iScript

http://www.bio-rad.c...A-Synthesis-Kit

and other forms of iScript:
http://www.bio-rad.c...iption-Reagents

Edited by memari, 20 March 2013 - 09:24 AM.

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Babak Memari

#5 Trof

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Posted 20 March 2013 - 01:55 PM

Roche Transcriptor also allows using either oligodT, random hexamer or both in reaction.
I'm not even sure if you need to buy kits, we were doing classic RT with separate components, so you can then buy what primers you want.

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