Re: [DIYbio] Thermal Cycler Based Bacterial Trasformation

Water bath is down for cleaning. This was just a trial run. The allure
of semi-automated transformation tempted me to try this. What may be a
solution to the aeration issue? Less time? Dispense the transformation
reaction into pre warmed LB flasks and shake for a while with no
antibiotics? Thanks for the insight!

Sent from my Windows Phone From: Tom Knight
Sent: 8/16/2013 4:45 PM
To: Sebastian Cocioba
Cc: Tom Knight
Subject: Re: [DIYbio] Thermal Cycler Based Bacterial Trasformation
Your outgrowth at 37 is a problem with no aeration. Otherwise, this
will likely work. Did you do a control transformation with a normal
heat shock?

On Aug 16, 2013, at 4:41 PM, Sebastian Cocioba 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.
>
>
> Sent from my Windows Phone
>
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