DNA concentration

Technical R&D your_name at cc.chiron.com
Thu Apr 20 16:30:45 EST 1995

In article <3n2dnr$lik at infoserv.rug.ac.be>, nater at gargamel.rug.ac.be
(Nancy Terryn) wrote:

> Dear netfriends
> can anybody help me to find a method that would allow me to quickly
measure the DNA concentration of many samples (say 500) I once heard of
some strips were you can spot DNA on but these seem to be rather
expensive. Somebody has an alternative? We use the PUC-vector if this can
help. Is it possible to do an assay using the LacZ gene?
> Any suggestions are wellcome..and will be reposted to the net for
general interest.
> --
> Nancy  

I have never done this myself, but I have heard that you can use agarose
gels with EtBr incorporated in it, then spot your DNA onto the gel and
compare its flourescence with a series of known standards. Make a normal
agarose gel with EtBR in it but pour the gel onto a 10 cm bacterial plate.
After it's hardended, drop <5 uL of your sample onto the gel and wait
until its diffused into it. Repeat this procedure with the standards (ex.
<5 uL each of 1 ug/uL, 0.5 ug/uL, 0.25 ug/uL, etc.). Place the plate over
a UV box and look to see how your unknowns compares with the series of
standards. The advantages: 1. cheap, 2. easy to prepare reagents, 3. Can
spot >30 samples onto a single plate (maybe even more) 4. sensitive down
to 50 ng of DNA, 5. relatively accurate.
Unfortunately I don't have all the details to the protocal, ie. what %
agarose, how thick should the gel be (should be pretty thin), maximum
volume to spot, etc. Hope this helps.


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