Electroporators for bacterial transformation

John Nash num208jn at MBDS.NRC.CA
Mon Nov 11 12:42:10 EST 1991


In article <1991Nov11.155502.1 at rsbs0.anu.edu.au>, jjw at rsbs0.anu.edu.au writes:
>John Nash writes:
>> One downside: the cuvettes aren't cheap and are not really usable..
>> from the top of my head, I think I pay something like $Cdn 2.50 -
>> $2.80 /cuvette.  Also, I find that one or two per box arc, which makes
>> a loud noise and scares the $%^& out of me for a few minutes.
>
>> John Nash                          |  Internet: Nash at biologysx.lan.nrc.ca
>
>We have a Biorad gene pulser and we're not too keen on paying vast amounts
>for new cuvettes.  Thus we reuse ours for all but the most VITAL experiments.
>We've been regularly reusing our EP cuvettes now for a few years.  No cross-
>contamination problems we've noticed, and we've got some cuvettes that have
>probably been used over 100 times.  Prior to your zap, add ~200ul of
>milliQ pure water and zap, then tip out and refill with your cells + DNA.
>After zapping, flush well 4-5 times with milliQ H20 and either store with 
>the cap on till your next use - or add the next batch of cells for zapping.
>The cuvettes should be eventualy retired, but you'll get lots of use from
>them before they start to arc.

Jeremy,

Thanks for the tip... the previous recycle tip I'd heard of involved
washing the cuvettes with bleach and then rinsing lots... I'm not too
keen on doing that.  Your tips sounds well worth trying.

Do you get much arcing?  I get at least one or two cuvettes per box
arcing, and I used to think that I had too much salt carried over from
my ligation (I just dilute ligations 1:1 with dH2O and zap 1 ul), BUT
repeating a sample with the same DNA which arced doesn't cause
rearcing, and sometimes DNA in dH2O arcs too.  I asked BioRad about
it, (and to summarize their response) they didn't have a clue, they
said that it "happens".

(BTW, we've met... I was doing my PhD at Monash Uni when I did a
sequencing course at RSBS in April 1983.  You showed me the sights of
Canberra -remember?  I AM a long way from home, now!).

cheers, 
John Nash                          |  Internet: Nash at biologysx.lan.nrc.ca
Institute for Biological Sciences  |   or: num208jn at mbds.nrc.ca 
National Research Council of Canada|  Ottawa,  Canada  K1A 0R6.	
==>  Disclaimer:  All opinions are mine, not NRC's!  <==



More information about the Methods mailing list