IGEM:MIT/2007/Notebook/2007-8-1

=Agenda=
 * 1) Decide on constitutive promoter for CPX
 * 2) *Transform promoter into DH5a
 * 3) *Plate transformants
 * 4) Check sequencing results for Mer/GFP 3A
 * 5) Perform flourescence assay on mer-gfp construct
 * 6) Clean out fridge

Flourescence Assay Protocol

 * 1) Draw sample of 1.5 ml from incubated cells with Hg
 * 2) Plot OD's
 * 3) Spin down 3 mins at 13000 rpm
 * 4) Aspirate and resuspend in 1.5 ml .9% NaCl
 * 5) Repeat 3
 * 6) Repeat 4
 * 7) Put 150ul in plate reader

Sequencing Results

 * mer.I13500 in pSB1AT3 came back really crappy. Biobrick prefix and suffix are partially present, cannot locate the sequence for GFP.
 * Need to send in the parts for sequencing.
 * Will run a colony PCR as a diagnostic?

Constitutive Promoter
Construct containing CPX and the polystyrene-binding peptide that will hopefully go into final system:

I14032/R0051.B0034.CPX-insert.

I14032 is a lacI repressable constitutive promoter. R0051 is a lambda cl repressable promoter.

Things That Got Cleaned Out
Ask Semmie.