To Do
- MOVE LAB
- Miniprep broth cultures
- Gel cultures
- Cryopreserve cultures
- Start assemblies(K235009, 13, 14), if we can come in on Sat, transform and plate as well.
- Test P1010 kan by transforming into DB3.1 and CCDB
8.30am
- Broth cultures removed from 37C water baths.
- Miniprep started.
- K235003 Cm A
- K235003 Cm B
- K235005 Cm
- K235010 Cm A
- K235010 Cm B
- J04450 Kan A 7A + IPTG
- J04450 Kan B 7A
- J04450 Kan A 7G + IPTG
- J04450 Kan B 7G
- P1010 Kan A
- P1010 Kan A (double for weighting)
- P1010 Kan B
- P1010 Amp A
- P1010 Amp B
- R0051 Amp A
- R0051 Amp B
- I0500 Kan A + IPTG
- I0500 Kan B
1.00pm
- Gels made. We ran out of ethidium bromide and got a new one with 10 mg/mL, so we had to dilute it with water to 2 mg/mL.
- Plates from yesterday show J04450 Kan 7A becomes much more red than 7G - to be expected, as 7G had an "inconsistent" sequencing, meaning mutations in the part sequence.
Gels started at 2.12pm
Gel 1:
| Contents |
4µL Ladder |
K235003 A |
K235003 B |
K235005 |
K235010 A |
K235010 B |
J04450 7A A (IPTG) |
| Banding? |
Yes |
Band |
Band |
Band |
Band |
Band |
Band |
Gel 2:
| Contents |
4µL Ladder |
J04450 7A B (no IPTG) |
J04450 7G A (IPTG) |
J04450 7G B (no IPTG) |
P1010 Kan A |
P1010 Kan A |
P1010 Kan B |
| Banding? |
Yes |
No |
No |
No |
Band |
Band |
Band |
Gel 3:
| Contents |
4µL Ladder |
P1010 Amp A |
P1010 Amp B |
R0051 A |
R0051 B |
I0500 A (IPTG) |
I0500 B (no IPTG) |
| Banding? |
Yes |
Band |
No |
No |
No |
Band |
No |
This shows that our assembled parts are likely working; we now have P1010 Kan and its substitute J04450; that R0051 seemed to have failed and that we have I0500.
(back to Lab notebook)
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