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

Agenda

1. Decide on constitutive promoter for CPX
   • Transform promoter into DH5a
   • Plate transformants
2. Check sequencing results for Mer/GFP 3A
3. Perform flourescence assay on mer-gfp construct
4. 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.