Protocol for RT-PCR in shrimp

jms93 jms93 at aber.ac.uk
Wed Mar 13 05:39:45 EST 1996


In article <4htus2$sdi at senior.nectec.or.th>, g3536459 at mucc.mahidol.ac.th 
says...
>
>        I have been working on the PCR for detection of Yellow-Head 
virus
>(YHV) in shrimps.  YHV is known as the RNA virus.  I used the Trizol
>Reagent (GIBCO BRL) for the isolation of RNA from hemolymph of shrimps. 
>The reagent, a mono-phasic solution of phenol and guanidine
>isothiocyanate, is an improvement of the single-step RNA isolation 
method
>developed by Chomczyniki and Sacchai.  Would anyone suggest any protocol
>that is more simple and rapid for RT-PCR in shrimp or any other 
organisms. 
>Thank you very much, 
>g3536459 at mucc.mahidol.ac.th
>
I'm not sure exactly what you are asking but as far as RNA isolation we 
use the RNeasy kit from Qiagen which produces good-quality RNA from as 
little as a gew mgs of tissue/cells which I have subsequently used 
succesfully in RT-PCR. For some samples, you do get DNA contamination so 
digestion with RNAse-free DNAse would be advised. The RT-PCR kit we use 
is from Promega and has been trouble-free since purchase.

Good luck

Jonathan Shillingford
jms93 at aber.ac.uk




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