roybina1 at yahoo.co.in
Wed Sep 25 08:18:24 EST 2002
I wish to know that how you can optimize the PCR conditions (eg. # of
cycles etc.) for a particular product you are amplifying. Do you have
to repeatedly run the PCR product in a gel and determine it.
My second question is that if I want to determine the best possible
forward and reverse primers for a particular gene (if I am unable to
find any published ones for that gene, nevertheless the sequence is
known), what are the good softwares for that. I am familiar with
Primer 3, but I want to recheck the primer sets again in a separate
program to prevent the possibility of primer-dimer formation. How am I
supposed to know if the chosen primers worked for me or not. By size
of the product after running on a gel?
A third question concers about choosing primers for a given set of
genetic markers. Do I have to plug in the marker (like D1S228) say in
PUBMED to know its sequence and then design a primer from there. Is
there any link that might help me to know if there are any primer sets
available as information.
Thank you all for helping me out.
More information about the Methods