DNA Reaction - Sequencing Single Stranded Templates
1. Aliquot DNA of the appropriate amount :
Asymmetric PCR Product 200 ng
Purified ss PCR Product 100 ng
M13 DNA 2 µg
Phagemid-induced ssDNA 2 µg
2. When ready to sequence, combine the Annealing Mix:
6 µL DNA, amount as above
2 µL 5X Sequenase Buffer
2 µL Primer, about 1 pmol, or 2-10 ng.
3. Pulse down in a microcentrifuge Tube.
4. Incubate at 70° for 5 min to completely unwind DNA strands
5. Slowly cool to 37° over 20 min to anneal primer.
6. Proceed with Sequenase DNA Sequencing Protocol.
Last Update: Thursday, 19-Jun-2014 11:50:23 PDT
This page has been viewed 195 times.
eckofoid at ucdavis.edu