Cloning - RE digestion clean up (Jan/06/2006 )
Usually I lost much DNA when phenol: Chloroform clean up. This time I want to use Kit. But most of the kits protocol written purificatin up to 10KB. Would you advice me which kit is good for large RE digested DNA. Restriction enzyme and DNA size are as follows.
1. PstI RE
2. Vector size 14Kb
3. Insert size 4Kb
Try using the geneclean system (Bio101) of the qiagen system
Could you just heat-inactivate your PstI? The lab next to mine does this all the time with their ligations and never has a problem.
If I remember correctly, 80 C for 20 minutes should do the trick (according to NEB). Don't quote me on that though!