Greetings LSOUTO:
Two questions for you-
1) How much formamide dye did you add to your dried samples? I add 2 - 3 ul
and load 1 ul.
2) If your control failed, what primer are you using?
I routinely freeze samples overnight with no adverse effects.
Make sure your plasmid concentration is correct and that your primer actually
binds to it.
Hope this helps.
Sheryl Christofferson
OMRF DNA Sequencing Facility
Oklahoma City, OK
LSOUTO wrote:
> Dear ABRF members:
> I need some advice from you:
> I ran some plasmid samples together with the PGEM control from PE, on a ABI
> 377.
> I use bigdyes;after cycle-sequencing samples were frozen and purified next
> day on centrisep columns. I evaporate on vaccuum centrifuge.
>
> I applied 1.5 ul of re-suspended samples, in a 24 comb.
> I use a 19.1 policarylamide solution
> Unfortunately, although I can see the signal of some sequence on the gel,
> the software makes no analysis "weak signal, analysis failed".
>
> What could possible be wrong??
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