PCR question

Shahram Mori smori at nmsu.edu
Fri Nov 11 23:03:05 EST 1994


Grad Student Lab 1 (gslab1 at unixg.ubc.ca) wrote:
:  Hi, there. Here is a question about PCR in my lab: if the amount of one 
: primer in the reaction is more than that of another one, let's say 30% more, 
: what will happen? Will the product mainly be a single strand headed with the 
: majority primer? For instance, if the majority primer is 1.1 kb far away from 
: the 5' end of its template, will the major product of this PCR be a 1.1 kb 
: DNA? 
: 	Any direction and comments are welcome.

: Please directly reply here or e-mail me at  xzhou at unixg.ubc.ca
First let's assume that your primers are highly gene specific. The amount
of primer that you add to a reaction is WAY hight generally. In fact most
people  wanna purify their PCR products away from the primers at the end of
30-40 cycles. Knowing the fact that your primers are present at saturating
amounts, I would expect that you would be left ( after 40 cycles) with
equal amounts of both strands. Your question really depends on the amount
of Template DNA, your [] of primers and the number of cycles that you are
carrying out.
Cheers

 --
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_ Saskatoon/SK/CANADA
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
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