EXOSAP-IT degrades my PCR products - (Jun/25/2008 )
I'm establishing the sequencing of 2 genes and experiencing some troubles. In the course of troubleshooting the reaction I've visalized 12 different ExoSAP purified PCR-products belonging to 2 different genes on a Agarose gel, realizing that almost all products (except of one of them) were shortened by up to 150 bp and/or heavily degraded after Exosap treatment. I've tried to use also an Exosap vial of our neighboor lab, getting the same results, so i don't think it's a matter of contamination. Interstingly 1 product out of twelve seems to be completely unaffected, while all other are in some way degraded and shortened. All product are 1000-1500 bp long and were amplified using the same Taq (Stratagene's Herculase II).
I would really appreciate if anybody here would have any hint for me, this thing is driving me really mad, since I've several hundreds of sequencing reactions to do!
Thank you very much for any help!
Hi goldideas! What du you mean with your reply? Did I miss something about ExoSAP-IT?
I'm having this same problem. Does anyone out there have any ideas?
Steve: Have you solved this problem yet?
Thanks for any thoughts.