Paul Shinn wrote:
>> OK, one more question about using water to resuspend samples. I've
> found when using only water, the initial peaks are VERY strong and then
> the signal quickly fades out. I started with the 50:50 of water and
> formamide/EDTA but the samples degrade by plate 4, however, the
> peaks were more even. For those of you running with water, is it just
> water or have you doped it with a touch of formamide or EDTA?
>
Paul, we changed our injection parameters to solve those problems.
modified parameters: injection voltage: 1500V
injection time: 20s
foil piercing: ON
flat field corrected: off
tank levels: OFF
Default parameters: run temp: 50'C
cuvette temp: 40'C
cap pressure: 1200psi
cap fill time: 1200s
max current: 800uA
pre-run voltage: 6000V
Pre-run time: 300s
sample volume: 2.5ul
wash volume: 20ul
run voltage: 5250V
sheath flow vol: 12000 counts
sheath flow period: 942msec
data delay time: 1500sec
run time: 10000sec
We are still using these modified paramters (except foil piercing is
off) for the water injection. This decreased the size of the initial
peaks and increased our readable run length significantly.
srlasky
--
Stephen R. Lasky, Ph.D. #
University of Washington #
Department of Molecular Biotechnology #
HSB K324 box 357730 #
Seattle, WA, 98195 USA #
email: srlasky at u.washington.edu #
Phone: 206-616-5865 #
Fax: 206-685-7301 #
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