-Did both the control and the plasmid samples fail, or did only the plasmid
samples fail??
if only the plasmid samples failed and the PGEM control samples worked
fine, then it sounds as if the concentration of the plasmid samples were
too low for the machine to detect the faint signal you see on the screen.
one way to avoid this is to spin down the solution of plasmid (before you
add the bigdye) in order to increase the concentration.
but if both PGEM and plasmid samples failed, then what was the amount of
PGEM solution used?
-susan
At 11:18 AM +0000 2/7/00, 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??
___________________________________________________________________
Susan K Fetics
Laboratory of MicroChemistry
Lindsley F. Kimball Research Institute
The New York Blood Center
310 E. 67th St., 3rd floor
New York, NY 10021
phone: (212) 570-3188
sfetics@nybc.org
For information about the MicroChemistry lab at NYBC or for request forms,
please visit:
http://www.nybloodcenter.org/framesets/FS-3C17.htm
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