bonhamp at duke.usask.ca (Peta C. Bonham-Smith) wrote:
>Has anyone been able to PCR amplify an insert directly from phage
>particles as oppposed to isolating phage DNA first? Do we need to treat
>the phage prior to PCR?
>University of Saskatchewan
>bonhamp at duke.usask.ca
We have been able to PCR directly from crude phae lysates.
We do so without any treatment and generally just take one microliter
of the lysate directly into the PCR tube. I have noticed that this
works fairly well if you have a high copy number of your target
template in the lysate, versus say, the contents of a genomic library
whereupon we get a high background...which may be explained by the way
the library was created...ie partial Sau 3A digest or just an over
abundance of nonspecific products. Our information was derived from
--Rover The Mad Molecular Biologist
rover at livnet.combarlow at picard.evms.edu