Biology-Online • View topic - nested pcr

Join for Free!
122508 members

nested pcr

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

Moderator: BioTeam

nested pcr

Postby 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.
Posts: 1
Joined: Fri Oct 02, 2009 10:19 am

Postby JackBean » Tue Oct 06, 2009 2:02 pm

Are you asking or just saying us?

Cis or trans? That's what matters.
User avatar
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Postby 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:

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)
User avatar
Inland Taipan
Inland Taipan
Posts: 3909
Joined: Thu Feb 03, 2005 2:46 pm
Location: Canada

Postby 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.
Death Adder
Death Adder
Posts: 81
Joined: Wed Apr 25, 2007 3:43 pm

Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 0 guests