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Team:UC Davis/Notebook/Week 2
From 2011.igem.org
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--Monday 6/20/11-- | --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: | + | Today we [[Team:UC_Davis/Protocols#nanodrop|nanodropped]] the 12 [[Team:UC_Davis/Protocols#digest|digested]] [[Team:UC_Davis/Protocols#errprone|mutated]] LacI promoters and the J61002 promoter screening plasmid such that we could [[Team:UC_Davis/Protocols#ligate|ligate]] them: |
{| border="1" style="color:white" | {| border="1" style="color:white" | ||
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| - | 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 then [[Team:UC_Davis/Protocols#ligate|ligated]] the 12 [[Team:UC_Davis/Protocols#errprone|mutated]] LacI promoters into the J61002 promoter screening plasmid. We had planned to [[Team:UC_Davis/Protocols#transformation|transform]] all of these [[Team:UC_Davis/Protocols#ligate|ligations]] but unfortunately ran out of LB+Carb plates. We were still able to [[Team:UC_Davis/Protocols#transformation|transform]] about 7 of the 12 [[Team:UC_Davis/Protocols#ligate|ligations]]. |
| - | We also rehydrated and transformed the B0015 terminator. | + | We also rehydrated and [[Team:UC_Davis/Protocols#transformation|transformed]] the B0015 terminator. |
| - | We miniprepped and digested the following parts: | + | We [[Team:UC_Davis/Protocols#miniprep|miniprepped]] and [[Team:UC_Davis/Protocols#digest|digested]] the following parts: |
* I13458 (with EcoRI and SpeI) | * I13458 (with EcoRI and SpeI) | ||
* I13453 (with EcoRI and XbaI) | * I13453 (with EcoRI and XbaI) | ||
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After yesterday's plate shortage, we took on the new task of making new LB+Carb plates. | 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: | + | We also [[Team:UC_Davis/Protocols#nanodrop|nanodropped]] many of the [[Team:UC_Davis/Protocols#miniprep|minipreps]] (MP) and [[Team:UC_Davis/Protocols#gel|gel purifications]] (GP) from earlier: |
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--Wednesday 6/22/11-- | --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. | + | [[Team:UC_Davis/Protocols#miniprep|Miniprepped]] B0015 and made [[Team:UC_Davis/Protocols#glycerol|glycerol stocks]] of I13453, I13458, and B0015. We also made some 10 mM IPTG from more concentrated stocks. |
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--Friday 6/24/11-- | --Friday 6/24/11-- | ||
| - | PCR purified B0034+E0240 and E0240. Digested B0034+E0240 with XbaI and PstI. | + | [[Team:UC_Davis/Protocols#pcrprep|PCR purified]] B0034+E0240 and E0240. [[Team:UC_Davis/Protocols#digest|Digested]] B0034+E0240 with XbaI and PstI. |
| - | Today we digested the following parts: | + | Today we [[Team:UC_Davis/Protocols#digest|digested]] the following parts: |
* B0034 (with SpeI and PstI) | * B0034 (with SpeI and PstI) | ||
* C0040 (with XbaI and PstI) | * C0040 (with XbaI and PstI) | ||
Revision as of 23:17, 15 September 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:
| 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)
