cloning PCR products

Michael Witty mw132 at mole.bio.cam.ac.uk
Fri Jan 25 07:47:28 EST 2002


On 25 Jan 2002, Purvi Bhatt wrote:

> Hi all,
> i too am facing a lot of problems with my 2kb insert which i am trying to
> clone into expression vector. I fail to understand why i get a  lot of
> colonies on vector self ligated plate while the test plate shows less
> colonies. also i am not getting even a single recombinant???? pl help as to
> where  can i be going wrong, is my vector not completely digested or what????
> troubleshooting needed. thanx.

Dear Purvi, post what you do and people can comment.  Do you gel purify
all your DNAs for example?  Do you use phosphatase?  What
restriction enzymes are you using to cut your PCR product?  Regards, Mike.




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