PCR directly from cell lines?
Nico van Belzen
belzen at hotmail.com
Thu Aug 19 08:43:11 EST 1999
I see no reason why it shouldn't work on cell lines. As you mention it can be
done on bacteria, that are harder to disrupt than eukaryotic cells. My advice is
to give it a try.
Nico van Belzen
Scott Currie wrote:
> Is there a method of amplifying a target directly from cells added to a PCR
> reaction? This can be performed on bacterial colonies and archival material
> such as parrafin blocks but I don't know if it can be performed on cells
> directly. I'd like to check my cell lines for stable transfectants or even
> transients) but at present do not have enough to extract genomic DNA or
> mRNA. This cell line has a habit of becoming resistant to G418 and wish to
> tackle this problem early instead of growing resistant non-transformed cells.
> If someone has another way of approaching this I'd be happy to listen.
> Additionally, I'm using pDNA3.1 and would like to know if only one
> construct integrates into the genome per cell. I'm told this is the case
> but cannot find evidence to support this (perhaps viral transfections are
> differrent). Is clonal populations determined by a southern, which
> indicates a single integrated construct, in a cloned cell line?
> I'm sorry this letter may seem naive, but thanks in advance,
> Scott A. Currie
> University of Melbourne
> Department of Surgery
> Western Hospital
> Footscray, Vic. 3011
> Tel: 61-3-9319
> Fax: 61-3-9317
> Email: s.currie at surgeryrmh.unimelb.edu.au
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