Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum

Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum (http://www.molecularstation.com/forum/)
-   Molecular Cloning Forum (http://www.molecularstation.com/forum/molecular-cloning-forum/)
-   -   Cloning bacteria: no growth (http://www.molecularstation.com/forum/molecular-cloning-forum/73672-cloning-bacteria-no-growth.html)

hianghao 08-27-2010 05:49 AM

Cloning bacteria: no growth
 
Hi

I am cloning competent E coli (tranformed with DNA ligated plasmid) on agar plate with ampicilin (50ug/mg) + X-gal/IPTG. There's no growth (blue/white) on my plate. Even when i use control plasmid with and without insert, there is no growth! What could be the problem?

bentelizabeth 10-01-2010 07:12 AM

Re: Cloning bacteria: no growth
 
1. It could be the competent cells have been thawed. Buy a new batch and test the control plasmid with them.

2. Your LB plates might contain too much antibiotic. Are you sure you added the right amount?

3. Find someone in your lab or building who has cloned successfully, then watch every step of what they do. If they use different volumes of cells to ligation, or a different waterbath, or different tubes (sometimes microfuge tubes can bind DNA), copy what they did and try again.

Good luck!


All times are GMT. The time now is 04:48 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved

Page generated in 0.08900 seconds with 11 queries