Exeter/30 August 2013

Exeter iGEM 2013 · Paint by Coli

Results from yesterday's transformations

ompF + RBS 100 µl equal amounts - 44 colonies

ompF + RBS 100 µl equal moles - 8 colonies

ompF + RBS 400 µl equal amounts - 107 colonies

ompF + RBS 400 µl equal moles - 35 colonies

[http://parts.igem.org/Part:BBa_J04450 RFP control] equal amounts - 15 colonies

[http://parts.igem.org/Part:BBa_J04450 RFP control] equal moles - 49 colonies

The ligation was carried out using two methods: equal volumes and equal moles. To calculate the equimolar volumes, we used the following data:

K592011, B0015, AMP plasmid, and RFP were from the digestions on the 29th. We pipetted the following volumes into four 0.6ml tubes:

New Part	A (Cyan + B0015, eq/vol)	B (RFP control, eq/vol)	C (Cyan + B0015, equimolar)	C(RFP Control, equimolar)
Plasmid	2 ul	2 ul	2 ul	2 ul
Cyan	2 ul	-	0.66	-
B0015	2 ul	-	-	-
B0015 (10x dilution)	-	-	1.2 ul	-
RFP	-	2 ul	-	0.74 ul
T4 ligase buffer	1.0 ul	1.0 ul	1.0 ul	1.0 ul
T4 DNA ligase	0.5 ul	0.5 ul	0.5 ul	0.5 ul
Non-nuclease water	2.5 ul	4.5 ul	4..64 ul	5.76 ul

The tubes were incubated at 16^oC for 30 minutes and then 80^oC for 20 minutes.

While carrying out the ligation, we ran a gel of our digested parts.

Take me back to the notebook.

Exeter iGEM 2013 · Paint by Coli