how to quantify pcr products

clarosa clarosa at biocomp.unl.edu
Tue Nov 7 18:28:51 EST 2000


Fluorimetry using Hoescht dye... or other is best.
http://lamarck.unl.edu/research/systematics/Orti/GO-lab/protocols/Fluorometer.html

We abandoned estimating by eye.. can also do spotting of dna on sarawrap ,
do a std curve, with ETBR stained DNA on the fluorimeter , an take a pic.

Karen Roberts wrote:

> I have tried using spec measurements but the yield is so low after
>
> I have purified the products that the readings give me next to or
> nothing.
>
> Next, I have run samples on a gel and dna is there but how can I
> quantify it?  I tried comparing the bands to bands of the marker (I
> called the company and got the formula for how much dna each marker band
> has based on how much you loaded of the marker) but I am getting numbers
> that make no sense when I do this gel estimation.
>
> I can give further details if you wish to email me privately but for
> now, what do others do to quantitate the amount of pcr product purified?
>
> MF






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