Fwd: What kits do people prefer for cDNA synth, and why?

---------- Forwarded message ----------
From: Nathan McCorkle <nmz787@gmail.com>
Date: Thu, Apr 15, 2010 at 1:35 PM
Subject: Re: What kits do people prefer for cDNA synth, and why?
To: diybio@googlegroups.com


The kit I used this morning included AMV reverse transcriptase, which has higher functioning temp which is better for denaturing secondary RNA structures... not sure how big a problem that is with mRNA.

It doesn't seem like RNase H activity is much of a problem since I'm looking to amplify only a certain mRNA, rather than make a library... does that make sense, or is RNase H activity just bad/useless ? 

On Thu, Apr 15, 2010 at 2:07 PM, Stanley Han <zhan528@gmail.com> wrote:

NEB kit doesn't seem to be robust. I don't know whay. Invitrogen has a Rt enzyme that has no endo H activity so that yo get more cDNA.Clontech seems to be good too. They are all more expensive.

On Thu, Apr 15, 2010 at 12:19 PM, Nathan McCorkle <nmz787@gmail.com> wrote:

I am looking for cDNA synth kits and settled on this one at $4.33 a reaction:

http://www.neb.com/nebecomm/products/productE6300.asp

I was settled actually, until I used a promega kit this morning after doing a trizol-chloroform RNA extraction. My professor said that trizol is cheap and yields high amounts of RNA, and said that he uses the promega kit because our school prefers to order through VWR... I am still waiting to hear if we get a discount through VWR or not.

If anyone has opinions about the kit I posted through NEB, or has a better recommendation please send quickly, as I'm trying to purchase stuff tomorrow, or at least have money set aside for its purchase. 

--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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Rochester Institute of Technology
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