Hi! I have a few questions:
1. Does too high a concentration of DNA hinder heat pulse transformation of XL1 Blue? If so, what is the optimum concentration?
2. I have a set of primers which doesn't seem to work. Do you know any website to check the feasibility of these primers? I know dimerization of primers, formation of hairpins and Tm affects the primer's efficiency but I don't know the exact guidelines for these criteria (acceptable # of self-dimerizing bp, optimum Tm, etc.).
Thanks a bunch!
Transformation and PCR questions
Started by bacchess, Jul 05 2004 03:59 PM
2 replies to this topic
#1
Posted 05 July 2004 - 03:59 PM
#2
Posted 05 July 2004 - 09:03 PM
Quote
Does too high a concentration of DNA hinder heat pulse transformation of XL1 Blue? If so, what is the optimum concentration?
So use no more than 1 ug DNA for a transformation reaction.
Quote
Do you know any website to check the feasibility of these primers?
Primer3 http://frodo.wi.mit....primer3_www.cgi
or check this page:
http://www.bioinform...520Design.shtml
#3
Posted 06 July 2004 - 07:35 PM
These threads may be of some help.
http://www.protocol-...p?showtopic=699
http://www.protocol-...?showtopic=2969
http://www.protocol-...p?showtopic=699
http://www.protocol-...?showtopic=2969













