"
Lab Sep 4 2009
From 2009.igem.org
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
