Help with Electroporation!!!
klenchin at macc.wisc.edu
Tue Apr 9 15:57:05 EST 1996
In article <pemanuel-0904960839110001 at beaker.ab.umd.edu>,
pemanuel at umabnet.ab.edu (Peter) wrote:
->My electroporation didn't work.
->I used the BIO RAD E coli gene pulser which lacks the ability to alter
->Their self test indicates the machine is OK.
Very graphical test for machine itself (hey, maybe their "self test isn't
working too!): put some salt solution in cuvette. If there is an arcing during
the pulse, then porator is OK.
->The cells are XL-Blue (300ul) at 2.5kV in a 0.2cm gap cuvette (Cold)
If this is true, it could be your problem.
Standard volume is 40-50 ul. With 300 ul the pulse is probably too short
to get decent efficiency.
->The plates are 100ug/ml carbenecillin.
->No colonies. nothing with my ligation mixes.
->Then I tried Supercoiled puC18. No colonies when I added 5 ul of a
->..01ng/ul solution of pUc18 which came from Stratagene.
->I even bought some Stratagene electro comp cells and they didn't work either.
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