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Team:TMU-Tokyo/Notebook/Protocol/
From 2012.igem.org
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<p class="description"> | <p class="description"> | ||
| - | <b>■Plasmid DNA Purification</b><Br> | + | <b>■Plasmid DNA Purification</b><Br></p> |
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■Genome DNA Purification</b>; Generation™ Capture Column (QIAGEN)<Br> | + | |
| + | <p class="description"> | ||
| + | <b>■Genome DNA Purification</b>; Generation™ Capture Column (QIAGEN)<Br></p> | ||
<Br> | <Br> | ||
| + | </p> | ||
| + | <p class="description3"> | ||
<b>1.</b> Add 200µL overnight cultures of E. coli in LB medium to the Capture Column.<Br> | <b>1.</b> Add 200µL overnight cultures of E. coli in LB medium to the Capture Column.<Br> | ||
<b>2.</b> Incubate 1 min at room temperature.<Br> | <b>2.</b> Incubate 1 min at room temperature.<Br> | ||
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<b>11.</b> Add 100µl DNA Elution Solution 2, incuvate 10 min at 99℃ with heat block.<Br> | <b>11.</b> Add 100µl DNA Elution Solution 2, incuvate 10 min at 99℃ with heat block.<Br> | ||
<b>12.</b> Centrifuge 20 s with tabletop centrifuge. Purified DNA will be released to solution.<Br> | <b>12.</b> Centrifuge 20 s with tabletop centrifuge. Purified DNA will be released to solution.<Br> | ||
| + | </p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■Restruction Enzyme Degestion</b><Br> | + | <p class="description"> |
| + | <b>■Restruction Enzyme Degestion</b><Br></p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■DNA Fragment Ligation</b><Br> | + | |
| + | <p class="description"> | ||
| + | <b>■DNA Fragment Ligation</b><Br></p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■Transformation</b><Br> | + | <p class="description"> |
| + | <b>■Transformation</b><Br></p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■Electrophoresis</b><Br> | + | <p class="description"> |
| + | <b>■Electrophoresis</b><Br></p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■LB Medium</b><Br> | + | <p class="description"> |
| + | <b>■LB Medium</b><Br></p> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
| - | <b>■PCR</b><Br> | + | <p class="description"> |
| + | <b>■PCR</b><Br></p> | ||
<Br> | <Br> | ||
Revision as of 20:12, 21 September 2012
■Experiment
1st week (8.13 - 8.19)
2nd week (8.20 - 8.26)
3rd week (8.27 - 9. 2)
4th week (9. 3 - 9. 9)
5th week (9.10 - 9.16)
6th week (9.17 - 9.23)
7th week (9.24 - 9.30)
■Protocols
Plasmid DNA Purification
Genome DNA Purification
Restruction Enzyme Degestion
DNA Fragment Ligation
Transformation
Electrophoresis
LB Medium
■Assay
Device1 Assay
Device2 Assay
Device3 Assay
Protocol
■Plasmid DNA Purification
■Genome DNA Purification; Generation™ Capture Column (QIAGEN)
1. Add 200µL overnight cultures of E. coli in LB medium to the Capture Column.
2. Incubate 1 min at room temperature.
3. Add 400µL DNA Purification Solution 1, incubate 1 min at room temperature.
4. Centrifuge 10 s with tabletop centrifuge.
5. Transfer the Capture Column to the Waste Collection Tube.
6. Add 400µL DNA Purification Solution 1, incubate 1 min at room temperature.
7. Centrifuge 10 s with tabletop centrifuge.
8. Add 200µl DNA Elution Solution 2. (Don't incubate.)
9. Centrifuge 10 s with tabletop centrifuge.
10. Transfer the Capture Column to DNA Collection Tube.
11. Add 100µl DNA Elution Solution 2, incuvate 10 min at 99℃ with heat block.
12. Centrifuge 20 s with tabletop centrifuge. Purified DNA will be released to solution.
■Restruction Enzyme Degestion
■DNA Fragment Ligation
■Transformation
■Electrophoresis
■LB Medium
■PCR
