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Template:Kyoto/Notebook/Aug 30
From 2013.igem.org
(Difference between revisions)
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==Aug 30== | ==Aug 30== | ||
===Gel Extraction=== | ===Gel Extraction=== | ||
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<div class="experiment"> | <div class="experiment"> | ||
<span class="author">No Name</span> | <span class="author">No Name</span> | ||
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</div> | </div> | ||
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===Miniprep=== | ===Miniprep=== | ||
<div class="experiment"> | <div class="experiment"> | ||
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</div> | </div> | ||
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===Colony PCR=== | ===Colony PCR=== | ||
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<div class="experiment"> | <div class="experiment"> | ||
<span class="author">No name</span> | <span class="author">No name</span> | ||
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</div> | </div> | ||
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===Ethanol Precipitation=== | ===Ethanol Precipitation=== | ||
<div class="experiment"> | <div class="experiment"> | ||
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===Restriction Enzyme Digestion=== | ===Restriction Enzyme Digestion=== | ||
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<div class="experiment"> | <div class="experiment"> | ||
| - | <span class="author">No | + | <span class="author">No name</span> |
{| class="wikitable" | {| class="wikitable" | ||
! ||8/30 RBS-lysis2 (9)||EcoRI||SpeI||10x bufferB||100x BSA||MilliQ||total | ! ||8/30 RBS-lysis2 (9)||EcoRI||SpeI||10x bufferB||100x BSA||MilliQ||total | ||
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|NC||0.5 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | |NC||0.5 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/22 pSB1C3-(2)||EcoRI||SpeI||10x Buffer||100x BSA||MilliQ||total | ! ||8/22 pSB1C3-(2)||EcoRI||SpeI||10x Buffer||100x BSA||MilliQ||total | ||
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|NC||1 µL||0 µL||0 µL||1 µL||0.1 µL||7.9 µL||10 µL | |NC||1 µL||0 µL||0 µL||1 µL||0.1 µL||7.9 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/22 pSB1C3-(2)||XbaI||PstI||BufferD||BSA||MilliQ||total | ! ||8/22 pSB1C3-(2)||XbaI||PstI||BufferD||BSA||MilliQ||total | ||
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|NC||1 µL||0 µL||0 µL||1 µL||0.1 µL||7.9 µL||10 µL | |NC||1 µL||0 µL||0 µL||1 µL||0.1 µL||7.9 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/21 tRNA-Spinach (1)||EcoRI||SpeI||10x BufferB||100x BSA||MilliQ||total | ! ||8/21 tRNA-Spinach (1)||EcoRI||SpeI||10x BufferB||100x BSA||MilliQ||total | ||
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|NC||0.5 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | |NC||0.5 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/21 tetR aptamer 12_1R (1)||EcoRI||SpeI||10xBufferB||100xBSA||MilliQ||total | ! ||8/21 tetR aptamer 12_1R (1)||EcoRI||SpeI||10xBufferB||100xBSA||MilliQ||total | ||
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|NC||0.4 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | |NC||0.4 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/21 pT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total | ! ||8/21 pT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total | ||
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|NC||0.5 µL||0 µL||0 µL||0 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | |NC||0.5 µL||0 µL||0 µL||0 µL||0 µL||0 µL||1 µL||0.1 µL||8.4 µL||10 µL | ||
|} | |} | ||
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{| class="wikitable" | {| class="wikitable" | ||
! ||8/21 pT181 antisense-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total | ! ||8/21 pT181 antisense-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total | ||
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|} | |} | ||
</div> | </div> | ||
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Revision as of 09:39, 22 September 2013
Contents |
Aug 30
Gel Extraction
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| PT181 atte(2) E+S | 305.3 | 2.2 | 1.66 | 0.36 |
| PT181 atte(2) X+P | 217.7 | 3.9 | 1.73 | 0.04 |
| PT181 anti(2) E+S | 347.7 | 3.1 | 1.88 | 0.31 |
| PT181 anti(2) X+P | 376.5 | 5.9 | 0.49 | 2.36 |
| PSBIC3 E+S | 436.6 | 30.7 | 1.03 | 1.28 |
| PSBIC3 X+P | 457.4 | 7.9 | 1.65 | 0.42 |
| Spinach E+S | 490.5 | 3.0 | 1.87 | 0.26 |
| tetR aptamer 12-1R E+S | 451.7 | -12.8 | 5.56 | -0.04 |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lysis2 9 | 264.6 | 1.97 | 2.21 |
| RBS-lysis2 12 | 220.0 | 1.89 | 2.46 |
| RBS-lysis2 14 | 195.3 | 1.88 | 1.53 |
Colony PCR
| Sample | base pair |
|---|---|
| 8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(8/20 LB Amp) (1)~(4) | 2143 |
| 8/29 Plux-RBS-GFP-DT(8/27 LB-CP) (1)~(2) | 1227 |
| 8/29 Pbad-larac-RBS-RFP{BBaI13516} (8/21 LB-CP) (1)~(2) | 2256 |
| 8/29 RBS-lysis3-DT-(1) | 1210 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 2min | 30cycle |
Ethanol Precipitation
| DNA |
|---|
| pT181 atte-(2) E+S |
| pT181 atte-(2) X+P |
| pT181 anti-(2) E+S |
| pT181 anti-(2) X+P |
| pSB1C3 E+S |
| pSB1C3 X+P |
| Spinach E+S |
| tetR aptamer 12_1R E+S |
Restriction Enzyme Digestion
| 8/30 RBS-lysis2 (9) | EcoRI | SpeI | 10x bufferB | 100x BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 7.5 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 18.2 µL | 30 µL |
| NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL |
| 8/22 pSB1C3-(2) | EcoRI | SpeI | 10x Buffer | 100x BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 8 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 17.7 µL | 30 µL |
| NC | 1 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL |
| 8/22 pSB1C3-(2) | XbaI | PstI | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 8 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 17.7 µL | 30 µL |
| NC | 1 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL |
| 8/21 tRNA-Spinach (1) | EcoRI | SpeI | 10x BufferB | 100x BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 4 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 21.9 µL | 30 µL |
| NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL |
| 8/21 tetR aptamer 12_1R (1) | EcoRI | SpeI | 10xBufferB | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 3 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 22.7 µL | 30 µL |
| NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL |
| 8/21 pT181 attenuator-(2) | EcoRI | SpeI | XbaI | PstI | 10x BufferB | 10x BufferD | 100x BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 4 µL | 0.5 µL | 0.5 µL | 0 µL | 0 µL | 3 µL | 0 µL | 0.3 µL | 21.7 µL | 30 µL |
| NC | 0.5 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0 µL | 0.1 µL | 8.4 µL | 10 µL |
| 2 cuts | 4 µL | 0 µL | 0 µL | 0.5 µL | 0.5 µL | 0 µL | 3 µL | 0.3 µL | 21.7 µL | 30 µL |
| NC | 0.5 µL | 0 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL |
| 8/21 pT181 antisense-(2) | EcoRI | SpeI | XbaI | PstI | 10x BufferB | 10x BufferD | 100x BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 9 µL | 0.5 µL | 0.5 µL | 0 µL | 0 µL | 3 µL | 0 µL | 0.3 µL | 16.7 µL | 30 µL |
| NC | 1 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0 µL | 0.1 µL | 7.9 µL | 10 µL |
| 2 cuts | 9 µL | 0 µL | 0 µL | 0.5 µL | 0.5 µL | 0 µL | 3 µL | 0.3 µL | 16.7 µL | 30 µL |
| NC | 1 µL | 0 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL |
