I net the ramp time from ice bath to 42C bath is way more than any old/cheap thermal cyclers.
I know its probably simple math to find a decent transitions time to change 50uL water from 4C to 42C using the heat capacity of a few gallons of water in a heat bath at 42C.
Simon, can you mathify us?
On Aug 16, 2013 1:42 PM, "Sebastian Cocioba" <scocioba@gmail.com> wrote:
-- --Anyone have any luck doing transformations using their PCR machine?
This is my protocol done on a 0.5mL Biometra UNO with no heated lid:
One fresh colony of HB101 per tube in 250uL of cold CaCl2 solution (100mM) + 1ng of Plasmid. All initial dispensing done cold onto prechilled PCR block holding at 4C.
Chill at 4C for 30mins
Heat at 42C for 45sec
Chill at 4C for 2min
Add 250uL LB Broth
Heat at 37C for 50mins
I get nothing. I'm using a simple biobricked gfp construct with AmpR. Would a PCRs relatively slow ramp time screw up the shock part of heat shock? My ramp time is about 1.8C per sec.
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