Cloning Pfu fragments...help|

John Cooper cooperj at nhlbi.nih.gov
Tue Nov 9 12:23:30 EST 1993


In article <1993Nov3.110034.19155 at gserv1.dl.ac.uk> GENETICA%IPCUCSC.earn at uk.ac.earn-relay ("L.Frigerio") writes:
>Path: nih-csl!lhc!biosci!daresbury!daresbury!news
>From: GENETICA%IPCUCSC.earn at uk.ac.earn-relay ("L.Frigerio")
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: Cloning Pfu fragments...help|
>Message-ID: <1993Nov3.110034.19155 at gserv1.dl.ac.uk>
>Date: 3 Nov 93 11:44:58 GMT
>Sender: list-admin at daresbury.ac.uk
>Distribution: bionet
>Lines: 12
>Precedence: first-class
>Original-To: "." <methods at uk.ac.daresbury>
>Hi net,
> 
>does anybody have experience with cloning of Pfu-amplified sequences?
>We tried to use standard blunt-end cloning and Klenow filling of the fragments
>but nothing seems to work...Any suggestions or informations will be greatly app
>reciated. Thanks in advance.
> 
>Lorenzo Frigerio
>Istituto di Genetica Vegetale
>UCSC Piacenza-Italy
>genetica at ipcucsc.bitnet
>botanica at imicilea.cilea.it


Try the strategene pCR-SK cloning kit. I use it for cloning these fragments.
Recombination is about 70% for white colonies-- good enough



More information about the Methods mailing list