FUNGAL cDNA

[Geneboy]

""Philip O'Brien"" <obrien at central.murdoch.edu.au> wrote in message
news:l03110703b862bb76f55b@[134.115.225.80]...
> Wha6t is the best method for extracting fungal RNA for making cDNA.  We
> have tried QIAGEN kits for extracting RNA and then subsequently purified
> mRNA.  However, although the control cDNA template provided with the
> Promega cDNA kit works fine, we do not get any product from the fungal
> mRNA.  Is there a preferred method for fungi?
>
> Phil

Hi Phil

Can't give a specific answer for fungi, I'm afraid, but I use Trizol (Gibco
BRL) for total RNA extraction and then SuperScript II (Invitrogen) for RT. I
don't isolate mRNA, and it seems to work fine for mammalian tissue samples
and cell lines. cDNA looks good, and I use this protocol to label cDNA for
microarrays.

Hope this helps

Chris

****************************************
Chris Cotsapas    [chrisc(at)student(dot)unsw.edu.au]
School of Biotechnology and Biomolecular Sciences
Faculty of Science
University of New South Wales
Sydney, Australia