Detection of annealed oligos - (Jan/05/2009 )
Hi there,
I am currently trying to insert/clone my annealed oligos (shRNA) into pSUPERIOR.neo vector (from Oligoengine). I was just wondering if you can detect or check if the oligos are annealed. Any suggestions? My oligos are 60-mer each with sticky ends.
Many thanks and have a nice day.
Regards,
Motz
-motz-
1. Are your oligos phosphorylated?
2. What is your annealing and ligation protocol?
Best,
Ginger
-Ginger Spice-
Yes you can by agarose gel electrophoresis. After annealing, run the annealed oligos along side equal amount of single stranded oligos, you will be able to see the ds-oligos migrate differently from ss-oligos.
-bioforum-
QUOTE (bioforum @ Jan 5 2009, 11:41 AM)
Yes you can by agarose gel electrophoresis. After annealing, run the annealed oligos along side equal amount of single stranded oligos, you will be able to see the ds-oligos migrate differently from ss-oligos.
I am thinking if you have more time and want to demonstrate convincingly to your supervisor, you should run non-denaturing acrylamide gel. Stain with etbr.
-TanyHark-