degenerate PCR - (Aug/18/2004 )

Hi,
I want to sequence my gene in different species. so I chose my degenerate primers and made my PCR, but I saw that just one primers can amplify my DNA. my primers are very unspecific, but I chose them in the very concerved part of my gene. I am sure that my gene is in this mix of amplification (lots of band) do you know a technic to extract my gene in this mix? or to have a more specific PCR...

-amira-

Hi, Amira,
Do you have a possibility to design primers for nested pcr? - it's a good way to fish out particular gene.

-meshuga-

QUOTE (meshuga @ Aug 18 2004, 12:04 PM)

Hi, Amira,
Do you have a possibility to design primers for nested pcr? - it's a good way to fish out particular gene.

I did it, but my nested primers are degenerate too, so I still have lots of bands with it.

-amira-