RNA isolation from frozen buffy coat

Jovita Castelino via methods%40net.bio.net (by J.M.Castelino from leeds.ac.uk)
Tue Aug 17 09:00:06 EST 2010


Dear Anita,

My name is Jovita Castelino and I am a PhD student at the University of Leeds. I saw your message on the bio.net forum, which was posted some years ago:

Hi,

could someone give me some suggestions how to increase the yield when
isolating total RNA from frozen buffy coat using the Trizol LS method?
Now I add Trizol LS to the buffy coat even before I start thawing it
because I am afraid RNases will work in the buffy coat. When thawed
the buffy coat/Trizol LS solution is difficult to pipette, very
viscous. I will be thankful for any help.

Anita


I wondered if you managed to solve the problem you had back then? If so, what did you do to get a better yield of RNA from frozen buffy coat? I am currently trying different kits to extract RNA from frozen buffy coat without much luck. I was thinking of trying out Trizol next but I want to make sure about it first. 

I would really appreicate any help/advice you could give me with this.

Thank you,

Jovita



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