nested pcr

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
shuk08
Garter
Garter
Posts: 1
Joined: Fri Oct 02, 2009 10:19 am

nested pcr

Post by shuk08 » Tue Oct 06, 2009 12:29 pm

How to get rid of larger non-specific bands appearing in 1st round of pcr just above the expected bands (of approx. 1.5 kb)? The non-specific bands from the 1st pcr are amplified in 2nd round of pcr using nested pair of primer.
Top
User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Tue Oct 06, 2009 2:02 pm

Are you asking or just saying us?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
Top
User avatar
canalon
Inland Taipan
Inland Taipan
Posts: 3909
Joined: Thu Feb 03, 2005 2:46 pm
Location: Canada

Post by canalon » Tue Oct 06, 2009 2:19 pm

I would try to optimize the first rund of PCR. Tips and tricks to do thta can be found here:
http://info.med.yale.edu/genetics/ward/tavi/Guide.html
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)
Top
roniadam
Death Adder
Death Adder
Posts: 81
Joined: Wed Apr 25, 2007 3:43 pm

Post by roniadam » Sat Oct 10, 2009 9:15 am

I had the same problem before. I advice you to use an adjuvant to improve your PCR effeciency. I used Q solution that comes with hotstar PCR master mix and it gives me a very good amplicon.
Top
Post Reply

Return to “Molecular Biology”

Who is online

Users browsing this forum: No registered users and 13 guests