Team:UC Davis/Notebook/Week 2
From 2011.igem.org
Week 2
--Monday 6/20/11--
Today we nanodropped the 12 digested mutated LacI promoters and the J61002 promoter screening plasmid such that we could ligate them:
Sample ID | ng/uL | 260/280 |
LacI 1 | 188.93 | 1.19 |
LacI 2 | 113.96 | 1.26 |
LacI 3 | 87.29 | 1.19 |
LacI 4 | 88.23 | 1.20 |
LacI 5 | 109.35 | 1.22 |
LacI 6 | 90.64 | 1.18 |
LacI 7 | 536.16 | 1.33 |
LacI 8 | 415.75 | 1.33 |
LacI 9 | 462.50 | 1.33 |
LacI 10 | 544.12 | 1.36 |
LacI 11 | 461.32 | 1.33 |
LacI 12 | 498.83 | 1.35 |
J61002 | 11.92 | 1.95 |
We then ligated the 12 mutated LacI promoters into the J61002 promoter screening plasmid. We had planned to transform all of these ligations but unfortunately ran out of LB+Carb plates. We were still able to transform about 7 of the 12 ligations. We also rehydrated and transformed the B0015 terminator.
We miniprepped and digested the following parts:
- I13458 (with EcoRI and SpeI)
- I13453 (with EcoRI and XbaI)
- R0010 (with SpeI and PstI)
- R0040 (with SpeI and PstI)
- R0051 (with SpeI and PstI)
--Tuesday 6/21/11--
After yesterday's plate shortage, we took on the new task of making new LB+Carb plates.
We also nanodropped many of the minipreps (MP) and gel purifications (GP) from earlier:
Sample ID | ng/uL | 260/280 |
J23101 MP | 338.75 | 1.87 |
I13456 MP | 245.92 | 1.86 |
I13453 MP | 181.48 | 1.85 |
R0010 MP | 115.98 | 1.86 |
R0051 MP | 101.21 | 1.89 |
E0040 MP | 240.71 | 1.82 |
R0040 MP | 80.18 | 1.92 |
C0012 MP | 165.16 | 1.87 |
R0010 GP | 22.42 | 1.73 |
I13458 GP | 16.63 | 1.86 |
R0040 GP | 20.39 | 1.62 |
I13453 GP | 15.10 | 1.74 |
R0051 GP | 7.27 | 2.69 |
--Wednesday 6/22/11--
Miniprepped B0015 and made glycerol stocks of I13453, I13458, and B0015. We also made some 10 mM IPTG from more concentrated stocks.
--Thursday 6/23/11--
After waiting most of the week some of our primers finally came in! We ran PCR with our B0034+E0240 forward primer and our E0240 forward primer.
--Friday 6/24/11--
PCR purified B0034+E0240 and E0240. Digested B0034+E0240 with XbaI and PstI.
Today we digested the following parts:
- B0034 (with SpeI and PstI)
- C0040 (with XbaI and PstI)
- C0051 (with XbaI and PstI)
- C0012 (with XbaI and PstI)
- J61002 (with EcoRI and PstI)
- psB1K3 (with EcoRI and PstI)