Pouring sequencing gels w/o leaking
Dennis J. Templeton
djt2 at po.CWRU.Edu
Fri Jul 9 14:37:02 EST 1993
In a previous article, jow at helix.nih.gov (Jim Owens) says:
>I wonder whether adding a small amount of non-ionic detergent, like
>0.01-0.05% NP-40, might be helpful for keeping bubbles under control.
>Once, when trying to sequence PCR fragments prepared from agarose gels, I
>was following a protocol that called for 0.05% NP-40 in the sequencing
>reaction. When the reactions were loaded on the gel the bubbles were so
>small that I could put the loading tip right on the gel surface without
>worrying about bubbles either disturbing the samples or staying in them.
>But on the other hand, there may be more bubbles formed in pouring the
>gel. It's just a thought I had.
I tried exactly this a few years ago. You have hit the problem right on
the head, any disturbance of the flow of solution onto the plate results
in a myriad of tiny bubbles that stream into the plate.
-just trying to save others from a mess like the one I made.
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