Cleaning seq. plates with "Bon Ami"??
Shiao Y. Wang
sywang at whale.st.usm.edu
Fri Apr 1 12:35:35 EST 1994
Dan Diaz (bl275 at cleveland.Freenet.Edu) wrote:
: In a previous article, balt at spieden.Stanford.EDU (Steven Balt) says:
: >Hello, all! Our sequencing gels have been pouring rather poorly
: >lately (no pun intended :) ) and we think we have narrowed down the
: >source of the problem to our new glass plates.
: >The glass plates have been scrubbed with "Bon Ami" brand cleanser
: >instead of the normal ICN "7X" cleaning solution or Windex. Has anyone
: >else experienced any weird phenomena after cleaning with Bon Ami??
: >(In our case, the acrylamide solution accumulates in the _center_ of the
: >glass plates instead of sliding down the sides, like it should.)
: here is the procedure we use in our lab. we use the BRL S2 apparatus with
: BRL plates and the IBI sharkstooth combs and spacers, since we dont like
: the IBI apparatus or the BRL combs and spacers.
: - soak brand new plates in 2M NaOH for 1-2 h and rinse thoroughly with
: deionized water. place a sliver of tape on one side of each plate to
: ensure consistent use of the same side of each plate for inside/outside.
: - clean both plates with glass plus followed by isopropanol
: - coat the small plate with Rain-X, a water repellent for windshields
: available at many auto parts stores. none of that yucky disgusting silane
: chloroform shit. put two coats of Rain-X on the small plate, let it
: dry, then wash again with glass plus and isopropanol
: - assemble the plate-spacer sandwich and clamp (tape is for sissies). we
: use three small clips on either side. the plate sandwich is laid
: horizontally on an empty pipet tip box or similar rectangular flat surface
: a few inches from the benchtop
: - prepare your gel mix and pour the gel horizontally, allowing the acryl
: mix to enter by capillary action. we put the mix in a plastic cylinder
: with lip, start on one corner of the gel, move to the other corner and then
: back to the middle, allowing the gel mix to move as a single front down the
: gel - its quick, easy and never fails. put some lab matting down to soak
: up possible spills.
: - put the comb in and clamp (we use two large clips placed so that they hold
: the comb in place, but dont let the clips extend beyond the bottom of the
: comb, i.e. clamp only where the clip is over comb or spacer, and never
: where there is only glass)
: - read a journal (a cool one) for 30-40 min and prepare your running buffer
: (or just prepare several litres of Tris-Taurine (glycerol-tolerant) buffer
: at once and store at RT)
: - use a thin spatula to separate the comb from the gel plate, then ease
: gently up. clean the surface of the gel plate which will contact the
: aluminum plate to ensure even heat distribution. make sure its dry.
: - set the gel in the apparatus and clamp tightly enough to avoid leaks, but
: not to maximum tightness. pour in running buffer and wash out the top of
: the gel with a pasteur pipet. start your prerun (we do 15-30 min at 40-60
: W when using Tris-Taurine)
: - get your sequenase reactions ready and wash yucky acrylamide from the
: comb with water. rinse the top of the gel and insert the comb slowly until
: the teeth just barely kiss the surface of the gel. tighten the gel until
: - rinse the wells once again and load. run the gel.
: - after the run, the plate treated with RainX will come off with minimal
: effort. we then rinse with water and set the plate to dry for next time.
: the larger plate goes into the fixing bath with the gel. once done, it
: also gets rinsed.
: - for the next run, simply clean with glass plus and isopropanol and youre
: ready to go. the RainX treatment will last for 20 gels or more. fun fun.
Thanks for the tip. One question: what is the purpose of soaking the new
glass plates in 2M NaOH? Won't the glass be etched or is that the intended
purpose so that acrylamide will stick better?
More information about the Methods