problem with sequencing in big dye v3
evetoulza at NOSPAMhotmail.com
Wed Oct 16 00:50:17 EST 2002
> Try to repurify your plasmids or change a kit for plasmid purification. If
it does not work, try to optimize conditions for PCR (change a temperature
for a denaturation step and an annealing). Be sure, that big dye kit is the
best option for your DNA.
We have already tried promega and qiagen minipreps kits, with same bad
results. In fact, not all the preps give bad results, between 20% and 80% of
them are nice, depends on what... whe don't know. Next week I will try to do
some PCR reactions in the lab with more initial denaturation time, more
cycles, and increased annealing temperature . Next minipreps will be eluted
in nuclease free water, and the PCR products will be purified with EDTA. I
really hope i could obtain significant upgrade in the quality of the
The plan to buy for the next time a big dye v3.1 (we use the 3.0).
thank you for your advices.
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