IGEM:MIT/2007/Notebook/2007-7-23

Agenda

1. Polystyrene Assay
   • Morning: get correct OD
   • Add AHL
   • Afternoon: Test
2. Digest I13500
3. Ligate 3A with I13500 and Mer in pSB1AT3 (so that, if we combine with polystyrene-binding plasmid into a single cell, can test for presence of both plasmids)
   • run overnight
4. dilute sequencing primers
   • sequence PHIE6P
   • sequence F2620+B0034+CPX
5. Order T7 antibody

Digestion of I13500

• Made two mixtures

4.5µl I13500(Miniprep #1, 223.5ng/µl)
1µl   XbaI
1µl   PstI
0.5µl BSA
5µl   NEB3 Buffer
38µl  H20
-------------------------
50µl  Total

• Placed in 37C @ 10:20am --> Take out @ 1:20pm

Diluted BioBrick Sequencing Primers

You can find the stock eppendorfs in the unnamed -20C fridge (Jason+Barry's room).

Diluted from 100µM stock to 32.2µM with total volume of 1.55mL each.

Made 1 VF2 and 1 VR for iGEM use.