Nylon membrane lot & false positive colony lift?
graham at biodec.wustl.edu
graham at biodec.wustl.edu
Tue Mar 25 07:14:51 EST 1997
What we have seen here recently suggests the possibility that different
postiviely charged nylon membrane lots (eg. Nytran, GeneScreen+, MSI
Magnacharge) might result in different amount of "background"
hybridization to E. coli library colony lifts. This is distinct from any
generic background darkening of the membrane itself -rather those clones
one would expect to be "negative" give different levels of signal
depending on the particular set of membranes prepared. Levels do appear
consitent within a particular batch prepared at once, but differ widely
when different lots of the same source of membrane are used.
I find it difficult to imagine how one charged nylon surface might
effect the hybridization of molecules bound to it differently than
another. However, different overall nucleic acid retention efficiencies
have been previously observed. Perhaps more proteins are bound by one
membrane lot which leads to a higher false signal with some membrane
lots?
Anyone seen it? If so, please e-mail me a copy of any responses
directly.
Thanks very much,
Jim
J. Graham PhD
Biology Department
Washington University of St. Louis
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