Dear Sequencers (and Genotypers),
A while back I asked for recommendations on the best way to purify BDT sequencing reactions for the PE 3700. Following are the recommendations I received:
For 96 well plates
Modified G50 Millipore method
For 384 well plates
Array-It from Telechem
Now I have one more question for more experienced PE 3700 users. How do you purify your formamide? There are two protocols from PE-ABI for purifying formamide, one using a mixed bed column method and one by the standard stirring/filtration with mixed bed resin. From our results with the 3700 it looks like the purity of the formamide is critical for good results. We can get over 650 base pairs of good sequence with the first run using HD Formamide from PE but later runs (reinjecting the same samples) show significant degradation in the length and quality of the sequences. Any tips would be greatly appreciated.
Best Regards,
Bob
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Robert B. Chadwick
Director, Genotyping/Sequencing Unit
Division of Human Cancer Genetics
Ohio State University
420 West 12th Avenue
Columbus, OH 43210
phone (614) 688-4783
FAX (614) 688-4761
e-mail chadwick-1 at medctr.osu.edu
web address http://gsu.med.ohio-state.edu/index.htm
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