5 Kb by RT-PCR

[Shahram Mori]

Pierre Commercon (pcomm at cismibm.univ-lyon1.fr) wrote:
: Hello

: I need help,
: How can be amplified by RT-PCR long fragment about 5Kb

: Thanks

: Pierre

: Pcomm at CISMIBM.univ-lyon1.fr
This is not really LONG PCR. I would just suggest using Taq and hot
starting your PCR. HoweverIf you have gene specific primers, I would suggest
using the rtTH ( from Perkin elmer ) and hot PCR.
This helps if your DNA has big time 2ndary structures as you can do the
annealing/extention at relatively high temps and help keep your DNA linear.
cheers,
 
-- 
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||