DNA quantitation in agarose/acrylamide gels

Jerry M. JM at spam.no.thanks
Fri Oct 2 10:07:20 EST 1998


In article <199809281413.WAA11830 at mail.shcnc.ac.cn>, shengyang at usa.net wrote:

> At 98-9-28 6:14:00, you wrote:
> >
> >Dear all,
> >This question was probably asked about 1000 times, however:
> >what is the most precise/fast/cheap technique to quantify intensity of
> >DNA bands in agarose/non-den. acrylamide gels (SYBR green, Ethidium
> >bromide etc.) or it is necessary to blot and hybridize the gel using
> >radioactive probe and Phosphoimager?
> 
> Hi, all:
> 
>         I think the fastest and cheapest way is to dyed with EB and
compare with the DNA molucule 
> size marker. I have no idea about what is the precised way :-(
> 
> Young

Another possibility, depending on your needs (and if it's possible for
your bug), would be to find a way to label the DNA with some isotope, like
with H3-dTTP. You would then calibrate a DNA concentration vs activity
curve for each labelling. After running the gel, the bands of interest
would be cutted and their activities determined.

I've done it with plasmid DNA, in an adenine auxotroph  mutant, using
H3-adenine. It's relatively fast, not extremely expensive and very
accurate.

Good luck,

   Jerry M.



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