In article <Pine.GSO.3.92.961008223124.10604A-100000 at chuma>, "Yi Zhang
(BIO)" <yzhang1 at chuma.cas.usf.edu> wrote:
> I am going to clone a large peice of viral DNA fragment into pPAC vector.
> This insert is 150 kb long. I have never done such a big piece of DNA
> cloning before. I would like to get help from anyone who has done such or
> who knows any literature (technical) on how to isolate this DNA from virus
This should be a useful reference, by the people who developed the PAC
Ioannou and de Jong, 1996
Construction of bacterial artificial chromosome libraries using the
modified P1 (PAC) system
In Current protocols in human genetics (ed. N.C. Dracopoli et al.), pp
5.15.1 - 5.15.24. John Wiley & Sons, New York
> without break the DNA, and what the best way is to transform the DNA into
> E. coli host (chemical competent cells or electroporation) .
Electroporation, for sure. DH10B is the preferred strain for PAC cloning.
Jose E. Mejia
Wellcome Trust Centre for Human Genetics
Windmill Road, Oxford OX3 7BN, UK
Tel: +44 1865 740021
Fax: +44 1865 742186
Email: jose.mejia at well.ox.ac.uk