ethanol treatment vs. plasmid loss
smori at nmsu.edu
Tue Nov 8 01:38:29 EST 1994
Luis Diaz (cascas at umich.edu) wrote:
: I have an mAb that recognizes an intracellular epitope of a T cell
: membrane glycoprotein. I want to clone the protein this mAb recognizes
: by transfecting a T cell cDNA library into NIH 3T3 cells and screening by
: flow cytommetry (FACS) with our FITC-labeled mAb. The problem I am
: concerned about is that to screen with my mAb we have to first
: permeabilize the transfected cells with 70% ethanol. I am worried that a
: successfully transfected plasmid would be lost after ethanol treatment.
: Am I wrong to assume the loss of a plasmid after 70% ethanol treatment or
: is this something that is likely to occur.
: Luis A. Diaz
: Medical Student
: University of Michigan
I would think that Permealizing doesn't mean total lysis of the cell. If 70%
ethanol doesn't lyse it, I would guess it should be alright.
Shahram Mori _/\_
Program in Molecular Biology _\ /_
Dept. of chemistry and Biochemistry Box 3C \_ _/
NMSU Las Cruces NM ||
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