Urgently need some help on a question for an assignment that has had me stumped for 2 days.
I'm supposed to clone and express a gene that encodes a protein/enzyme known as CobT produced by the bacterium Mycobacterium avium paratuberculosis. Now I need to design a primer to help me amplify, clone and express the protein. Problem is I'm totally confused on how to go about this.
How do you design a primer? I know this is a super basic question but the multitude of tutorials I've seen have me all confused.
I tried using the ncbi primer blast tool but it seemed all the primers it gave me were inside my target sequence which makes no sense, how do you use the ncbi tool to pick primers?
Is there a difference between a primer for cloning and one for PCR?
Any help would be appreciated.
The CobT nucleotide sequence is attached below, the target sequence is highlighted in blue.
Edited by taiju, 12 April 2013 - 04:59 AM.