mspeek at ebc.ee
Fri Dec 11 04:10:23 EST 1992
wetsel_r at wums.wustl.edu wrote:
: Greetings netters:
: This is probably a sequencing FAQ but here goes: I have a technician in the
: lab that can do a "single" load reasonably well. The problem is when the
: run is extended or even "double" loaded. What we're getting is fuzzy
: , sometimes doubled bands which are difficult to decipher. Is the Urea going
: off, temp too low/high, any suggestions would be appreciated...
: OH! BTW - I should note that this is double-stranded sequencing with oligos
: and using Sequenase.
: Thanks in advance,
: haviland at kids.wustl.edu
I have noticed the same phenomenon when using commercial SEQ
apparatus (Macrophor, LKB), but never with home-made machines and
gels run with contact with metal plate. So, I propose this is
some kind of combination of heat and high voltage (thermostbilized
plates) that makes longer runs difficult or impossible to read.
Indeed, when reducing voltage (down to 1 kv) bands appear to be
sharper. I hope this explanation helps you somehow.
Mart (mspeek at ebc.ee)(ESTONIA)
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