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Template:Kyoto/Notebook/Sep 22
From 2013.igem.org
(Difference between revisions)
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===Gel Extraction=== | ===Gel Extraction=== | ||
<div class="experiment"> | <div class="experiment"> | ||
| - | <span class="author"> | + | <span class="author">No name</span> |
{| class="wikitable" | {| class="wikitable" | ||
!Lane||DNA||Enzyme | !Lane||DNA||Enzyme | ||
| Line 60: | Line 60: | ||
|1||100bp ladder||-- | |1||100bp ladder||-- | ||
|- | |- | ||
| - | | | + | |2~4||Pcon-pz12-1R-DT(E-1A)||-- |
|- | |- | ||
| - | | | + | |6~8||Pcon-spinach-DT(E-1A)||-- |
|- | |- | ||
| - | | | + | |10~12||Pcon-attenuator PT181(E-1A)||-- |
| - | |- | + | |
| - | + | ||
|} | |} | ||
| - | |||
| - | |||
| - | |||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! | + | !Lane||DNA||Enzyme |
| + | |- | ||
| + | |1||100bp ladder||-- | ||
| + | |- | ||
| + | |2~4||DT(1-3)||-- | ||
|- | |- | ||
| - | | | + | |6~8||aptamer12-1RDT(1-3)||-- |
|- | |- | ||
| + | |10~12||Pcon-PT181 attenuator(1-5A)||-- | ||
|} | |} | ||
| - | |||
===digestion=== | ===digestion=== | ||
Revision as of 05:49, 26 September 2013
Contents |
Sep 22
Colony PCR
| Sample | base pair |
|---|---|
| 9/20 Plac-PT181 attenuator(1~4) | 664 |
| 9/20 Plac-PT181 antisense(1~6) | 369 |
| 9/20 Pcon-PT181 attenuator-aptamer-DT(1~6) | 859 |
| 9/20 Pcon-PT181 antisense-spinach-DT(1~8) | 723 |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 9/21 spinach-DT | 197.9 | 1.67 | 1.54 |
| 9/21 Plux | 125.9 | 1.71 | 1.27 |
| 9/21 Plux-RBS-GFP-DT-(1) | 168.0 | 1.62 | 1.73 |
| 9/21 Pcon-PT181 attenuator-(1) | 368.3 | 1.57 | 1.62 |
| 9/21 RBS-GFP-DT | 532.8 | 1.75 | 1.78 |
| 9/21 Pcon-RBS-GFP-DT | 845.2 | 2.02 | 1.70 |
Colony PCR
| Sample | base pair |
|---|---|
| 9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1~2) | 283 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 2min10s | 30cycles |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2~4 | Pcon-pz12-1R-DT(E-1A) | -- |
| 6~8 | Pcon-spinach-DT(E-1A) | -- |
| 10~12 | Pcon-attenuator PT181(E-1A) | -- |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2~4 | DT(1-3) | -- |
| 6~8 | aptamer12-1RDT(1-3) | -- |
| 10~12 | Pcon-PT181 attenuator(1-5A) | -- |
digestion
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 3 | RBS-lacZα-DT | XbaI&PstI |
| 4 | RBS-lacZα-DT | XbaI&PstI |
| 5 | RBS-lacZα-DT | XbaI&PstI |
| 6 | RBS-lacZα-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(XbaI&PstI) | 9.8 | 1.50 | 0.39 |
Colony PCR
| Sample | base pair |
|---|---|
| pSB1C3+P-KaiBC① | 283 |
| pSB1C3+P-KaiBC② | 283 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 2min | 30s | 30s | 18s | 30cycles |
Colony PCR
| Sample | base pair |
|---|---|
| pSB1C3+P-KaiBC① | 283 |
| pSB1C3+P-KaiBC② | 283 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 2min | 30s | 30s | 18s | 30cycles |
Colony PCR
| Sample | base pair |
|---|---|
| pSB1C3+P-KaiBC① | 283 |
| pSB1C3+P-KaiBC② | 283 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 2min | 30s | 30s | 18s | 30cycles |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | 9/18 Plac (SpeI & PstI) | 3.0 µL | 9/18 RBS-lysis3-DT (XbaI & PstI) | 12.4 µL | 3.5 µL |
| experiment | 9/12 Ptet-pT181 antisense | 1.0 µL | 9/18 spinach-DT | 11.9 µL | 3.5 µL |
| experiment | 9/13 Pcon-RBS-luxR-DT | 2.1 µL | 9/10 Plux-RBS-GFP-DT | 5.6 µL | 3.5 µL |
| experiment | 9/13 Pcon-pT181 attenuator (SpeI & PstI) | 2.0 µL | 9/18 aptamer 12-1R-DT (XbaI & PstI) | 6.3 µL | 3.5 µL |
| experiment | 9/18 Plac ②(SpeI & PstI) | 3.0 µL | 9/18 aptamer 12-1R-DT(XbaI & PstI) | 4.5µL | x3.5 µL |
Liquid Culture
| Sample | medium |
|---|---|
| 9/2 spinach-DT-2 | -- |
| 8/16 Plux-4 | -- |
| 8/27 Plux-RBS-GFP-DT-1 | -- |
| 9/5 Pcon-pT181 attenuator-1 | -- |
digestion
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 3 | RBS-lacZα-DT | XbaI&PstI |
| 4 | RBS-lacZα-DT | XbaI&PstI |
| 5 | RBS-lacZα-DT | XbaI&PstI |
| 6 | RBS-lacZα-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(XbaI&PstI) | 9.8 | 1.50 | 0.39 |
Transformation
| Name | Sample | Competent Cells | Plate |
|---|---|---|---|
| 9/17 pSB1C3-Pcon-pT181 attenuator-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-aptamer 12_1R-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-RBS-tetR-DT x2 | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-PkaiBC | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-RpaB | 2 µL | 20 µL | CP |
| 9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-pT181 attenuator(XbaI&PstI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-spinach-DT | 2 µL | 20 µL | CP |
| 9/14 RBS-GFP-DT-PkaiBC | 2 µL | 20 µL | CP |
| Plux-RBS-lysis3-DT | 2 µL | 20 µL | CP |
| 9/20 Ptet-pT181 antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/20 Pcon-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 Plac-RBS-lysis3-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer 12_1R | 2 µL | 20 µL | Amp |
| 9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2 µL | 20 µL | Amp |
| Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20Pcon-pT181attenuator-aptamer-DT | 2 µL | 20 µL | Amp |
| 9/17 Plac-pT181-attenuatorx2 | 2 µL | 20 µL | Amp |
| 9/15 Pcon-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| 9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| PUC19 | 2 µL | 20 µL | Amp |
PCR
| Sample | base pair |
|---|---|
| pT181 attenuator(pSB1C3) | -- |
| Pcon-RBS-lacZα-DT | -- |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94 °C | 94 °C | 55 °C | 68 °C | -- |
| 5 min | 30 s | 30 s | 3 min | 30 cycles |
Transformation
| Name | Sample | Competent Cells | Plate |
|---|---|---|---|
| 9/17 pSB1C3-Pcon-pT181 attenuator-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-aptamer 12_1R-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-RBS-tetR-DT x2 | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-PkaiBC | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-RpaB | 2 µL | 20 µL | CP |
| 9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-pT181 attenuator(XbaI&PstI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-spinach-DT | 2 µL | 20 µL | CP |
| 9/14 RBS-GFP-DT-PkaiBC | 2 µL | 20 µL | CP |
| Plux-RBS-lysis3-DT | 2 µL | 20 µL | CP |
| 9/20 Ptet-pT181 antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/20 Pcon-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 Plac-RBS-lysis3-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer 12_1R | 2 µL | 20 µL | Amp |
| 9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2 µL | 20 µL | Amp |
| Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20Pcon-pT181attenuator-aptamer-DT | 2 µL | 20 µL | Amp |
| 9/17 Plac-pT181-attenuatorx2 | 2 µL | 20 µL | Amp |
| 9/15 Pcon-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| 9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| PUC19 | 2 µL | 20 µL | Amp |
