Hello, I have a solution wih a DNA fragment with a sticky-end cut. It is possible ligate the oligo to this extreme without anneal it with the complementary?
Thnak you
Hi, my opinon the DNA and oligo must have complementary sticky end that mean it must cutting in the same restriction enzymes. good luck
if the sticky ends are, for instance, CC at one end and GG at the other in the DSDNA then an oligo with end bases CC will anneal to the GG end but not to the CC end of the fragment
You can do it with T4 RNA ligase. Don't be fooled by the name, it works on ssDNA as well. Go to NEB's site for details.
DNA ligase T4 can involve to this kind of reaction without any problem,The additional use of T4 RNA ligase can increase the effectiveness of the reaction. An increase in efficacy can be achieved without T4 RNA ligase by adding to the reaction mixture PEG6000 and NaCl.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology