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Team:TU Delft/16 September 2010 content
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 |
