DNA quantitation method wanted

Jeff crowther at rschp1.anu.edu.au
Wed Mar 9 01:41:31 EST 1994

In article <3253.2d73f345 at mbcl.rutgers.edu>, marian at mbcl.rutgers.edu wrote:
>     I am looking for a quick and effiecient method for quantitation
> of DNA.  I want to use the method to control for cell number in
> cell cultures.  I need most of the cell layer, so I only have a
> small percentage of the sample availabe for quantitation.  I am
> aware of Invitrogen's "sticks", but these are not very cost-efficient.
> I am condsidering Maniatis' protocol for etbr quantitation of
> droplets on saran wrap, but have serious reservations about the 
> relaibility of this method.  Any suggestions?

I have used the Maniatis method for quantitation of DNA in protein preps.
Rather than using saran wrap I use a 1% agarose mould of 10x5 4mm wells.
The DNA/EtBr in appropriate amounts are mixed in small ed's and then put
wells. Run a set of std's from known DNA conc on the gel block.
DNA drops are of uniform size due to wells as are std's
No/low background from gel
Low diffusion at 1% over a moderate length of time
I find quantitation is not too bad from a reasonable number of stds but
depends on the level of precision you want
If you want to get carried away Scan gel/photo and do densitometry on it.I
have not done this so don't know how accurate it is.

Hope it helps. If you have any further queries e-mail me
crowther at rsc3.anu.edu.au

Jeff C.
Centre for Molecular Structure and Function
Research School of Chemistry
Australian National University

Who knows what the signature says so many people use my computer

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