blunt end ligation problems
Posted 17 February 2005 - 08:05 AM
Basically, I did like follows:
1. Diagest the plasmid with Hinc II.
2. Gel purify the fragment I want.
3. Do ligation and transform into DH5a
However, I tried several times and just could not get any clone.
What is the problem? Is there anything wrong with my idea?
Posted 17 February 2005 - 09:14 AM
Posted 17 February 2005 - 11:04 AM
Posted 20 February 2005 - 11:53 AM
Usually, when you buy competent cells, they come with a control plasmid. or you can use any other plasmid that you know works.
Posted 20 February 2005 - 11:56 AM
Posted 20 February 2005 - 08:25 PM
It may sound silly and simple but sometimes it does the trick.
I may question the activity of the ligase, but I would try changing the water first.
If that doesn't work, then I am out of ideas, my friend.
Posted 21 February 2005 - 06:55 PM