Plasmid isolation problem

[manoj oak]

Dear Grains,
   
  I cloned LMW glutenin genes (of wheat) in pMOS-T blue and also pGEM-T cloning vectors, I confirmed positive clones using colony PCR, I used vector primers and also specific LMW glutenin primers. I got expected size amplicon.  Then I incubated positive clones to isolate plasmids (alkaly lysis method) in large ammount for sequencing. But i am not getting the plasmids. 
   
  Please let me know what is going wrong in my case. 
   
  I appreciate your help in this regards.
   
  with best regards
  sincerely yours
   


If you judge people, you have no time to love them. -- Mother Teresa

Manoj D Oak Ph. D.
Scientist 
Genetics Department
Plant Science Division 
Agharkar Research Institute
Agarkar Road 
Pune 411 004 
INDIA 
Phone +91-20-25653680 (Off) 
E-mail: manojoak at yahoo.com
 		
---------------------------------
Stay in the know. Pulse on the new Yahoo.com.  Check it out. 
-------------- next part --------------
An HTML attachment was scrubbed...
URL: <http://brie4.cshl.edu/pipermail/gramene/attachments/20060829/892a2424/attachment.html>