AMOUNT OF PRIMER IN DNA FINGERPRINTING

[Shiao Y. Wang]

GORDON BETTS (betts at ORION.ETSU.EDU) wrote:
: I am about to order some primer for an undergraduate fingerprinting exercise
: I am trying to duplicate and need to know how much primer to use.  I will
: use the D1S80 primer.  The procedure by Budowle et al. (Am. J. Hum. Genet.
: 48:137) calls for 1 uM.  Isn't this a lot?  When I look at order forms, they
: often have a checkbox for 50 or 100 nM.  Lot of difference!  

I think there is a slight confusion here. I have never done fingerprinting
but have done a good bit of PCR; 1 uM is 1 pmol of primer per ul. When you
order primers, the checkbox is for 50 or 100 nmole (and not nM). For a
20-mer, 50 nmole is roughly 310 ug of DNA and dissolving it in one ml of
water would give you a 50 uM stock.

I hope this helps and please correct me if anyone spots a mistake.

Shiao Wang
University of Southern Mississippi
sywang at whale.st.usm.edu