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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-