Scale up of ligation reaction.

Rodney Earl Pettway earlpettway at netscape.net
Thu May 18 13:51:05 EST 2000


Thanks again for your help!!!!!!!!!!!

Have a great day!

"Hiranya S. Roychowdhury" wrote:

> With the right conditions you can scale up ligation reactions to 50uL. I
> have had good success in that volume using the 5x ligation buffer containing
> 25% PEG8000. Indeed, while trying to clone some larger genomic fragments,
> that was the volume that seemed to work best for me. Also, the i:v ratio
> needs to be kept at 1:3 to 1:5 with larger vector sizes. If you have access
> to the Sambrook et al. manual, go through the chapter therein on cosmid
> cloning. You will get a very comprehensive idea about the conditions required.
>
> At 01:14 PM 5/18/00 -0500, Rodney Earl Pettway wrote:
> >I was wondering if it is possible to scale up a cosmid/genomic DNA
> >ligation reaction or is it better to make several small reactions.
> >I know that I post a lot of questions to this news group but I am trying
> >a lot of new things and I want to get the advice of the experts before I
> >try it myself.
> >
> >thanks!!!!
> >
> >Rodney Earl Pettway
> >USDA -ARS Cereal Disease Lab
> >University of Minnesota
> >
> >
>
> Dr. Hiranya Sankar Roychowdhury
> GENE LAB/ EPPWS
> New Mexico State University
> Las Cruces, NM 88003
> Ph. (505) 646-5785
> hroychow at nmsu.edu
>
> ---
-------------- next part --------------
A non-text attachment was scrubbed...
Name: earlpettway.vcf
Type: text/x-vcard
Size: 248 bytes
Desc: Card for Rodney Earl Pettway
Url : http://iubio.bio.indiana.edu/bionet/mm/methods/attachments/20000518/c698c9e8/earlpettway.bin


More information about the Methods mailing list