Nested PCR
Moderators: honeev, Leonid, amiradm, BioTeam
Nested PCR
Can you use one of the primers you have used in the first round of your nested PCR in the second round along with a new primer situated within the first round product???
Nested PCR means that two pairs of PCR primers were used for a single locus.
The selection of alternative and similar primer binding sites gives a selection of products, only one containing the intended sequence. The product from the first reaction undergoes a second run with the second set of primers.
It is very unlikely that any of the unwanted PCR products contain binding sites for both the new primers, ensuring the product from the second PCR has little contamination from unwanted products of primer mis-binding and alternative primer target sequences.
The selection of alternative and similar primer binding sites gives a selection of products, only one containing the intended sequence. The product from the first reaction undergoes a second run with the second set of primers.
It is very unlikely that any of the unwanted PCR products contain binding sites for both the new primers, ensuring the product from the second PCR has little contamination from unwanted products of primer mis-binding and alternative primer target sequences.
DIP JYOTI CHAKRABORTY.
NATIONAL CHILD SCIENTIST(2004).
RESEARCH WORKER ON WETLAND. MEMBER OF RAMSAR; GLAND, SWITZERLAND. MEMBER OF EPILEPSY FOUNDATION; CONNECTICUT.
NATIONAL CHILD SCIENTIST(2004).
RESEARCH WORKER ON WETLAND. MEMBER OF RAMSAR; GLAND, SWITZERLAND. MEMBER OF EPILEPSY FOUNDATION; CONNECTICUT.
Who is online
Users browsing this forum: No registered users and 16 guests