Template:Kyoto/Notebook/Aug 24
From 2013.igem.org
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|8/22 pT181 antisense (EcoRI & SpeI)||8||1.13||0.82 | |8/22 pT181 antisense (EcoRI & SpeI)||8||1.13||0.82 | ||
|- | |- | ||
- | |8/22 Fusion3m2 attenuator (XbaI & PstI)||5||2. | + | |8/22 Fusion3m2 attenuator (XbaI & PstI)||5||2.27||1.50 |
|- | |- | ||
|8/22 Fusion3m2 attenuator (EcoRI & SpeI)||34||1.16||1.02 | |8/22 Fusion3m2 attenuator (EcoRI & SpeI)||34||1.16||1.02 | ||
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===Restriction Enzyme Digestion=== | ===Restriction Enzyme Digestion=== | ||
- | |||
<div class="experiment"> | <div class="experiment"> | ||
<span class="author">No name</span> | <span class="author">No name</span> | ||
{| class="wikitable" | {| class="wikitable" | ||
- | !tetR aptamer 12_P(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total | + | ! ||tetR aptamer 12_P(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total |
|- | |- | ||
- | |2.7µL||0.5µL||0.5µL||3µL||0.3µL||23µL||30µL | + | |2 cuts||2.7µL||0.5µL||0.5µL||3µL||0.3µL||23µL||30µL |
|- | |- | ||
- | |0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | + | |NC||0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL |
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
- | !tetR aptamer 12_1R(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total | + | ! ||tetR aptamer 12_1R(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total |
|- | |- | ||
- | |2.6µL||0.5µL||0.5µL||3µL||0.3µL||23.1µL||30µL | + | |2 cuts||2.6µL||0.5µL||0.5µL||3µL||0.3µL||23.1µL||30µL |
|- | |- | ||
- | |0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | + | |NC||0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL |
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
- | !tetR aptamer 12_M(1)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total | + | ! ||tetR aptamer 12_M(1)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total |
|- | |- | ||
- | |2.7µL||0.5µL||0.5µL||3µL||0.3µL||23µL||30µL | + | |2 cuts||2.7µL||0.5µL||0.5µL||3µL||0.3µL||23µL||30µL |
|- | |- | ||
- | |0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | + | |NC||0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL |
|} | |} | ||
+ | </div> | ||
===Master Plate=== | ===Master Plate=== | ||
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===Electrophoresis=== | ===Electrophoresis=== | ||
- | |||
<div class="experiment"> | <div class="experiment"> | ||
<span class="author">No name</span> | <span class="author">No name</span> | ||
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!Lane||Sample||Enzyme1||Enzyme2 | !Lane||Sample||Enzyme1||Enzyme2 | ||
|- | |- | ||
- | |1|| | + | |1||100bp ladder||-||- |
|- | |- | ||
|2||tetR aptamer 12_P||EcoRI||SpeI | |2||tetR aptamer 12_P||EcoRI||SpeI | ||
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|7||tetR aptamer 12_1M||-||- | |7||tetR aptamer 12_1M||-||- | ||
|- | |- | ||
- | |8|| | + | |8||100bp ladder||-||- |
- | |} | + | |}<br> |
+ | [[File:Igku Aug24 Electrophoresis(N1)-pic.jpg]]<br> | ||
+ | <br> | ||
+ | |||
{| class="wikitable" | {| class="wikitable" | ||
!Lane||Sample||Enzyme1||Enzyme2 | !Lane||Sample||Enzyme1||Enzyme2 | ||
|- | |- | ||
- | |1|| | + | |1||100bp ladder||-||- |
|- | |- | ||
|2||tetR aptamer 12_P||EcoRI||SpeI | |2||tetR aptamer 12_P||EcoRI||SpeI | ||
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|4||tetR aptamer 12_1M||EcoRI||SpeI | |4||tetR aptamer 12_1M||EcoRI||SpeI | ||
|- | |- | ||
- | |5|| | + | |5||100bp ladder||-||- |
|} | |} | ||
+ | <br> | ||
+ | [[File:Igku Aug24 Electrophoresis(N2)-pic2.jpg]]<br> | ||
+ | |||
+ | <br> | ||
</div> | </div> | ||
- |
Latest revision as of 16:18, 25 September 2013
Contents |
Aug 24
Gel Extraction
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
8/22 aptamer 12_P (EcoRI & SpeI) | 88 | 21.3 | -- |
8/22 aptamer 12_1R (EcoRI & SpeI) | 63 | 23.1 | -- |
8/22 aptamer 12_1M (EcoRI & SpeI) | 40 | 22.0 | -- |
8/23 Fusion6 antisense (EcoRI & SpeI) | 6.0 | 1.60 | 0.17 |
8/23 Fusion6 antisense (XbaI & PstI) | 6.0 | 1.71 | 0.25 |
8/23 pT181 antisense (EcoRI & SpeI) | 6.0 | 2.22 | 0.15 |
8/23 pT181 antisense (XbaI & PstI) | 6.0 | 2.10 | 0.04 |
8/22 Plac (SpeI & PstI) | 6.0 | 1.47 | 0.09 |
8/22 Fusion1 antisense (EcoRI & SpeI) | 10 | 2.16 | 0.01 |
8/22 J23100 (SpeI & PstI) | 6.0 | 1.52 | 0.35 |
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
8/22 Spinach (EcoRI & SpeI) | 8.0 | 1.15 | 0.77 |
8/22 Fusion6 antisense (XbaI & PstI) | 44 | 1.07 | 0.96 |
8/22 Fusion1 antisense (XbaI & PstI) | 2 | -- | 1.00 |
8/22 pT181 antisense (XbaI & PstI) | 3. | 2.01 | 0.40 |
8/22 pT181 antisense (EcoRI & SpeI) | 8 | 1.13 | 0.82 |
8/22 Fusion3m2 attenuator (XbaI & PstI) | 5 | 2.27 | 1.50 |
8/22 Fusion3m2 attenuator (EcoRI & SpeI) | 34 | 1.16 | 1.02 |
8/22 Fusion1 attenuator (XbaI & PstI) | 14 | 1.40 | 0.62 |
8/22 Fusion1 attenuator (EcoRI & SpeI) | 16 | 1.27 | 0.61 |
8/22 pT181 attenuator (XbaI & PstI) | 5 | -- | 0.88 |
- These samples were performed ethanol ethanl precipitation.
Restriction Enzyme Digestion
tetR aptamer 12_P(2) | EcoRI | SpeI | 10x Buffer B | 100x BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.7µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 23µL | 30µL |
NC | 0.3µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
tetR aptamer 12_1R(2) | EcoRI | SpeI | 10x Buffer B | 100x BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.6µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 23.1µL | 30µL |
NC | 0.3µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
tetR aptamer 12_M(1) | EcoRI | SpeI | 10x Buffer B | 100x BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.7µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 23µL | 30µL |
NC | 0.3µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
Master Plate
Number | Use LB plate(+Kan) |
---|---|
1 | entA- #1 |
2 | entA- #1 |
3 | entA- #1 |
4 | entA- #2 |
5 | entA- #2 |
6 | entA- #2 |
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | 100bp ladder | - | - |
2 | tetR aptamer 12_P | EcoRI | SpeI |
3 | tetR aptamer 12_P | - | - |
4 | tetR aptamer 12_1R | EcoRI | SpeI |
5 | tetR aptamer 12_1R | - | - |
6 | tetR aptamer 12_1M | EcoRI | SpeI |
7 | tetR aptamer 12_1M | - | - |
8 | 100bp ladder | - | - |
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | 100bp ladder | - | - |
2 | tetR aptamer 12_P | EcoRI | SpeI |
3 | tetR aptamer 12_1R | EcoRI | SpeI |
4 | tetR aptamer 12_1M | EcoRI | SpeI |
5 | 100bp ladder | - | - |