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Team:Kyoto/Notebook/DMS Synthesis
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アメフラシノート
7/31
Restriction Enzyme Digestion
| Sample | Volume | EcoR1 | Spe1 | Buffer | MilliQ | TOTAL |
|---|---|---|---|---|---|---|
| 3/24 PCR(dddD)cloning | 3.2 | 1 | 1 | 3(H) | 21.8 | 30 |
| pSB1C3 | 18.5 | 1 | 1 | 3(H) | 6.5 | 30 |
| pSB1C3(control) | 6.2 | 0 | 0 | 3(H) | 2.8 | 10 |
8/1
Gel Extraction
| Lane | DNA |
|---|---|
| 1 | 1kbp ladder |
| 2 | 3/24 PCR(dddD)cloning E,S |
| 3 | 3/24 PCR(dddD)cloning E,S |
| 4 | Blank |
| 5 | pSB1C3 |
| 6 | pSB1C3 |
| 7 | Blank |
| 8 | 1kbp ladder |
| 9 | 3/24 PCR(dddD)cloning(control) |
| 10 | pSB1C3(control) |
8/1
Ligation
| Vector | inserter | 10xt4LigaseBuffer | t4Ligase | MilliQ | TOTAL |
|---|---|---|---|---|---|
| 1.5 µL(8/1 pSB1C3 2047bp 13.3µg/mL 20μg) | 6.9 µL(8/1 dddD 2545bp 10.8µg/mL 74.60µg) | 2µL | 1µL | 8.6µL | 20µL |
8/1
transformation
| Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Plate |
|---|---|---|---|---|
| pSB1C3 | 3 | 30 | 165 | SOC(132µl) |
8/3
Electrophoresis for confirmation
| Lane | Sample |
|---|---|
| 1 | pSB1C3 dddD 1 |
| 2 | pSB1C3 dddD 2 |
| 3 | pSB1C3 dddD 3 |
| 4 | 1kb ladder(3µL) |
| 5 | 1kb ladder(1µL) |
8/3
Gel Extraction
| Lane | DNA |
|---|---|
| 1 | 1kbp ladder |
| 2 | Blank |
| 3 | pSB1C3 25µμL 2070bp |
| 4 | Blank |
| 5 | pSB1C3 5µL |
| 6 | Blank |
| 7 | 8/2 cloning dddD 25µL 2544bp |
| 8 | Blank |
| 9 | 3/24 PCR(dddD)cloning(control) |
| 10 | 8/2 cloning dddD 5µL |
