Lab Sep 4 2009

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.

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