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Team:UC Davis/Notebook/Week 2
From 2011.igem.org
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== Week 2== | == Week 2== | ||
--Monday 6/20/11-- | --Monday 6/20/11-- | ||
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* R0040 (with SpeI and PstI) | * R0040 (with SpeI and PstI) | ||
* R0051 (with SpeI and PstI) | * R0051 (with SpeI and PstI) | ||
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--Tuesday 6/21/11-- | --Tuesday 6/21/11-- | ||
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<img src="https://static.igem.org/mediawiki/2011/b/bb/Pouring.JPG" width="639" height="428"> | <img src="https://static.igem.org/mediawiki/2011/b/bb/Pouring.JPG" width="639" height="428"> | ||
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--Wednesday 6/22/11-- | --Wednesday 6/22/11-- | ||
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--Thursday 6/23/11-- | --Thursday 6/23/11-- | ||
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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. | 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. | ||
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--Friday 6/24/11-- | --Friday 6/24/11-- | ||
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* J61002 (with EcoRI and PstI) | * J61002 (with EcoRI and PstI) | ||
* psB1K3 (with EcoRI and PstI) | * psB1K3 (with EcoRI and PstI) | ||
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Revision as of 22:59, 22 August 2011
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== 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:
{| border="1"
|'''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:
{| border="1"
|'''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
|}
</div>
</div>
--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)
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