DNA Reaction - Sequencing Double Stranded Templates - Nonidet Method
1. Purify DNA and resuspend in TE to a concentration of 100 ng/mL.
2. Prepare Denaturing Mix:
1.0 µL DNA (approximately 100 ng of a 1 kb PCR product; 1-5 mg plasmid)
2.0 µL 1 µM Oligonucleotide (about 2 pmol)
1.0 µL 5% Nonidet P-40 in water
1.0 µL ddH2O
3. Incubate at 100° for 3 min. Immediately place on dry ice for 5 min.
4. Thaw during a 15 sec spin and transfer to ice
5. Continue with Sequenase sequencing, using this mixture as the Annealing Mix.
Last Update: Thursday, 19-Jun-2014 11:50:22 PDT
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