kliman@mbcl.rutgers.edu (04/11/91)
We have found that repeated silanization of glass plates used for DNA
sequencing gels eventually leads to problems in pouring the gel, i.e., the gel
mix does not flow evenly and forms persistent bubbles. While we can correct
this by overnight soaking in 10-20% NaOH, we would rather not leave large
volumes of NaOH out overnight on a regular basis. And, of course, we would
prefer to decrease our exposure to silane..
Has anyone else experienced similar problems? We'd appreciate any
suggested alternatives to silanization that could be used on a daily basis.
Thanks. NUM208JN@NRCCAD.NRC.CA (John Nash) (04/11/91)
>To: methods-and-reagents@ocelot.rutgers.edu >From: kliman@ocelot.rutgers.edu >Subject: Seeking alternatives to silanizing gel plates >Date: 10 Apr 91 22:03:25 GMT > We have found that repeated silanization of glass plates used for DNA >sequencing gels eventually leads to problems in pouring the gel, i.e., the gel >mix does not flow evenly and forms persistent bubbles. While we can correct >this by overnight soaking in 10-20% NaOH, we would rather not leave large >volumes of NaOH out overnight on a regular basis. And, of course, we would >prefer to decrease our exposure to silane.. We use Aquasil from Pierce Chemicals. It is a water-soluble, silicon-based "glass coater" for hospital equipment. I dilute 10 ml in 2.5 litres of water and do all the glassware I can find. You "rinse" the glass in it, wash in water, then either bake for 1 hour or let it sit at RT for a day, then wash. This stuff actually makes sequencing gels easier to pour. It lasts for about 5 to 10 gels. When you fix the first gel run after washing the plates, the plate in the fixer loses some of its coating. This is handy for subsequent sequencing gels, as the gel tends to come off onto that plate each time. This prevents half the gel sticking to one plate and half the gel sticking to the other. Hope this helps. cheers, John. (Bitnet: NUM208JN@NRCCAD.NRC.CA)
roca@psl.wisc.edu (Alberto I. Roca) (04/11/91)
In article <384.2803536d@mbcl.rutgers.edu>, kliman@mbcl.rutgers.edu writes: > > We have found that repeated silanization of glass plates used for DNA >sequencing gels eventually leads to problems in pouring the gel, i.e., the gel >mix does not flow evenly and forms persistent bubbles. While we can correct >this by overnight soaking in 10-20% NaOH, we would rather not leave large >volumes of NaOH out overnight on a regular basis. And, of course, we would >prefer to decrease our exposure to silane.. > Has anyone else experienced similar problems? We'd appreciate any >suggested alternatives to silanization that could be used on a daily basis. >Thanks. We coat one side of one plate with Rain-X (available from hardware stores). Other people spray their plates with Pam. In either case, it is unnecessary to treat with silanization. ================================================================= Alberto I. Roca Internet: roca@vms.macc.wisc.edu Biochemistry Bitnet: roca@wiscmacc 420 Henry Mall University of Wisconsin-Madison Madison, Wisconsin 53706 USA
kim@m44.unm.edu (04/16/91)
>In article <384.2803536d@mbcl.rutgers.edu>, kliman@mbcl.rutgers.edu writes: > (stuff omitted) >We coat one side of one plate with Rain-X (available from hardware stores). >Other people spray their plates with Pam. In either case, it is unnecessary >to treat with silanization. > I like the idea of cheap household products that can do the same functions as expensive "biotechnology" stuff. I have been told, as part of the oral tradition of the lab where I work, that glassware and plasticware to be used in handling RNA should be silanized to prevent adherence and 'hold-up' of nucleic acids. This is applied to micropipet tips, micro-tubes, and columns for poly(A)+ selection. Does anyone have any idea why Rain-X cannot be used for this purpose? I imagine that Rain-X out of an unopened bottle is as free of RNAases as other factory-fresh reagents, or could treated surfaces be autoclaved for good measure? Daniel Kim