Sequencing plates problem

coryell at nauvax.ucc.nau.edu coryell at nauvax.ucc.nau.edu
Thu Jan 19 11:54:31 EST 1995


Another sequencer, here...  Additional information: "silanize" probably
is (or possibly is) gamma-methacryloxypropyltrimethoxy silane (from
Sigma, catalog number M-6514); at least this is what we have used.
Mix 9 ml ethanol, 150 ul 10% acetic acid, and 15 ul gamma-methacryloxy-
propyltrimethoxy silane.  Pour onto the plate you want the gel to stick
to, spread across the plate with gloved hands (in a hood is good), let dry
15 minutes, then wash the plate with distilled water and ethanol to remove
excess "sticky" silane.  One coating lasts several uses.  We had no problem
with floating gels; we even agitated (rotary shaker) during gel fixing
with no problem.  Our gels were 0.2mm-1.0mm exponential.  Getting the gel
off was another question.  We soaked the gel in 10%NaOH for a day or more,
although another sequencer said (this is only heresay, mind you!) that
soaking in water works as well.
        The side you DON'T want to stick can be coated with Repel-Silane
from LKB (it's somewhat expensive) or Rain-X (for car windshields, in the
automotive supplies at your local Wal-Mart or hardware/auto parts store).
I have had good results with both.  Good luck.



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