PCR ?

David Micklem dmicklem at cmgm.nospam.invalid
Sat Jul 15 21:44:52 EST 2000


In article <301C7F169C2 at scammonden.shef.ac.uk>, David Keszenman-Pereyra
<D.K-Pereyra at sheffield.ac.uk> wrote:

>I want to integrate a selectable cassette randomly into the genome. 
>Then I want to sequence the flanking regions of the integration site.
>Is there any PCR method to do it quickly ????? (single primer 
>amplification ?????)
>

This is done pretty routinely in the fly world - see
<http://www.fruitfly.org/p_disrupt/inverse_pcr.html>
for a protocol for inverse-PCR off the ends of P-element transposons. I
think the Berkeley Drosophila Genome Project do/did this on a large
scale to identify STS's. I've followed their protocol on small numbers
of lines with a high success rate.

David

-- 
D.R. Micklem,          Time flies like an arrow... Fruit flies like a banana. 
Beckman Center,        Email:dmicklem at cmgm.stanford.edu
Stanford University    Phone: +1 (650) 723-6650
Stanford, Ca 94305       Fax: +1 (650) 725-6044
USA                    Unsolicited email will incur a US$100 processing charge.






More information about the Methods mailing list