IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-27

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Miniprep of LC1PUR1 and LC2PUR1 transformants
We miniprepped the transformants we inoculated yesterday. There are 8 total. We've recorded their concentrations below (we dropped the "PUR1" from the ends of their names).

All DNA was eluted in 50 µL H2O.

PCR mutagenesis #6
The main purpose of this PCR is to determine the optimal concentration of template. The smearing in our previous PCRs suggests that we've been using too much template (or our primers are contaminated).

We've chosen to use LC1 3kb #1 and LC2 3kb #1 as our samples.


 * Or however much template is normally used in the positive control

A black dot on the vial indicates the LC1 set (done by Jeff), a purple dot indicates LC2 (done by David). Hetmann did the primer controls.

Note: I didn't include negative controls with template but no primer or Vent, since I think those controls should wait until we discover the appropriate concentration of template to use.

Note: we ran out of the Cat primer, so Jeff's positive control probably won't work.

Note: we made a calculation error; our master mix had 25% more water than it's supposed to. Hopefully this won't affect our results (each reaction is still the same total volume, so the concentrations of template and primer were not affected).