Team:UC Davis/Notebook/Week 2

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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 |}
--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. </div>

--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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