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From 2010.igem.org

Emulsifier

Yesterdays cultures grown over night. Since Top10 cells activate Lac promoters by themselves, I did a quick and dirty experiment on those cultures.

1 ml culture was spin down for 2 min at max speed.

The supernatant was used in the emulsification assay.

#	Description	OD600	absorption (493 nm)	absorption (493 nm) with Sudan II
1	Top 10 (control)	2.177	2.302
2	Top 10 with 206	1.703	1.862
3	Sup Top 10 (control)		0.013	0.341
4	Sup Top 10 with 206		0.015	0.349

So the conclusion is that there is no difference between the control and the transformed cultures. So we just have to wait until we have the K12 strain.

Alkane degradation

009A and 010A were grown up overnight, and isolated using a mini-prep. The plasmids were digested and checked on a gel (cut and uncut). The gel looked good - no 'disappeared' plasmids as yesterday.

#	BioBrick	Cut with	Buffer	BSA	Expected Length (bp)	Correct?
0	Smartladder (5μl)
1	009	uncut				Yes
2	009	SpeI, PstI	Buffer 2 (NEB)	BSA (1x)		Yes
3	010	uncut				Yes
4	010	XbaI, PstI	Buffer 2 (NEB)	BSA (1x)		Yes
5	018	uncut
6	018	EcoRI, SpeI	Buffer 2 (NEB)	BSA (1x)		Yes
7	019	Uncut				Yes
8	019	EcoRI, XbaI	Buffer 2 (NEB)	BSA (1x)		Yes
9	001	uncut				Yes
10	005	Uncut				Yes
11	021AK	uncut				Yes

The digestion products were ligated, and commercial TOP10 cells were transformed with the ligation mixtures.

BioBrick	Plasmid BB	Insert	Product
028A	009A	010A	J61100-rubA4-J61100-rubR
026A	018A	019A	J61101-ADH-J61107-ALDH