what makes electroporation cuvette "bang"? - (Nov/29/2005 )
During electroporation, sometimes when i add the voltage up to 2500v for 2.5mm cuvette, it will bang.I know there are multiple factors for that, could someone tell me exactly what makes electroporation "bang"? my lab uses Bio-Rad electroporator.
The problem is low resistance, due to excess salts in the cuvette. Dialyze the DNA against water by floating a Millipore VSWP 0025 membrane, shiny side up, on the surface of water in a petri dish. Pipet the ligation mix onto the membrane surface, cover the petri dish, and let it sit for 1/2 hour. Pipet the solution off and mix with cells, then electroporate.
Some common causes:
* Too much salt in the DNA (do as phage suggested or just use less DNA).
* Too much salt in the cells, left-over from the growth medium. Dilute cells in ice-cold MQ water, pellet at 4 degrees, then gently resuspend again in ice-cold MQ water.
* A human sweat / condensation mixture on the outside of the cuvette - so wear gloves and dry the cuvette thoroughly (but quickly) immediately prior to zapping it.
the bang comes from salt...as you've read on the previous posts.
before doing an electroperation, clean the ligation solution. i ethanol precipitate the solution before hand, others in my lab use quiagen colums to clean it up.
be careful with using bacteria that goes bang. they, unfortunately, get fried during the process, and might not work.
Thx for all above.i have gotten some tips from ur answers.when the voltage are high,the electrodes discharge, so i think if there were not salts in the solution,it would bang naturally.why doesn't the electroporator bang in ”normal“ electroporation？maybe it has become a physical question.confused....
just a side note, I would still plate out Electroporation samples that arc or 'go bang'. I have had success many times with 'bang' samples.
yes,sometimes i will work although the microbe gets fried someone told me that the'bang' sample might bring mutants.now i wonder whether it is suitable to use the bang ones.hope for more interpretation....
you can try pelleting the legation with glycogen