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Team:Kyoto/Notebook/DMS Synthesis
From 2014.igem.org
(Difference between revisions)
| Line 12: | Line 12: | ||
<span class="author">Nakamura Matsumoto</span> | <span class="author">Nakamura Matsumoto</span> | ||
<table class="note"> | <table class="note"> | ||
| - | <tr><th>Sample</th><th>Volume</th><th>EcoR1</th><th>Spe1</th><th> | + | <tr><th>Sample</th><th>Volume</th><th>EcoR1</th><th>Spe1</th><th>Buffer</th><th>MilliQ</th><th>TOTAL</tr> |
<tr><td>3/24 PCR(dddD)cloning</td><td>3.2</td><td>1</td><td>1</td><td>3(H)</td><td>21.8</td><td>30</td></tr> | <tr><td>3/24 PCR(dddD)cloning</td><td>3.2</td><td>1</td><td>1</td><td>3(H)</td><td>21.8</td><td>30</td></tr> | ||
<tr><td>pSB1C3</td><td>18.5</td><td>1</td><td>1</td><td>3(H)</td><td>6.5</td><td>30</td></tr> | <tr><td>pSB1C3</td><td>18.5</td><td>1</td><td>1</td><td>3(H)</td><td>6.5</td><td>30</td></tr> | ||
| Line 34: | Line 34: | ||
<tr><td>10</td><td>pSB1C3(control)</td></tr> | <tr><td>10</td><td>pSB1C3(control)</td></tr> | ||
</table> | </table> | ||
| + | |||
| + | <h2>8/1</h2> | ||
| + | <h3>Ligation</h3> | ||
| + | <span class="author">Yasuda</span> | ||
| + | <table class="note"> | ||
| + | <tr><th>Vector</th><th>inserter</th><th>10xt4LigaseBuffer</th><th>t4Ligase</th><th>MilliQ</th><th>TOTAL</tr> | ||
| + | <tr><td>1.5 μL </td><td>6.9 μL </td><td>2 μL </td><td>1 μL </td><td>8.6 μL </td><td>20 μL </td></tr> | ||
| + | </table> | ||
| + | |||
<h2>8/3</h2> | <h2>8/3</h2> | ||
Revision as of 02:44, 10 September 2014
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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 |
8/31
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 | 6.9 μL | 2 μL | 1 μL | 8.6 μL | 20 μL |
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 25μL 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 |
