Error Prone PCR

olivier Namy namy at infobiogen.fr
Tue Jun 6 14:08:56 EST 2000


hello,

I have a protocole to do error prone PCR which works very well. I have been
randomly mutagenised a 2Kb gene with this protocol (the number of mutations
is beetwen one and five per fragment)
I used :
7mM MgCl2
0,5mM Mncl2
1mM d(C/T)TP
0,2mM d(A/G)TP
2,5 Units of taq polymerase
10uM each oligonucleotides.
(Becarfull the choice of buffer is very very important, in my hand onle CETUS
(from Perkin Elme) buffer works.
the elongation time is lowest in these conditions approxymatively 1.5min per
Kb).

If you want more explanations, you can email me to namy at infobiogen.fr

Olivier Namy.






More information about the Methods mailing list