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 ||
88003
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