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.
Sent from my Windows Phone
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.
Sent from my Windows Phone
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