Sequencing off Bacterial Genomic DNA with ABI Big Dye Terminators
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The following protocol and cycle conditions courtesy of Cheryl Heiner-ABI
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Protocol:
Big Dye Terminator Miundefined 16μl
ThermoFidelasundefined_1μl
Primer (18-24mer)15-30pmoles
Genomic DNA Template 3-6μg
Final Reaction Volume 40μl
Cycle Conditions:
95 degC- 5 min.initial hold
45 cycles of
95 degC- 30 sec.
55 degC- 20 sec.
60 degC- 4 min.
4 degC hold
Purify through G-50 spin columns and load under standard gel conditions
Description: TF,KIT BTD RR-1000
