Team:Chiba/jk/β
From 2008.igem.org
(Difference between revisions)
Line 581: | Line 581: | ||
③ -> OK ・・・30ng? | ③ -> OK ・・・30ng? | ||
|} | |} | ||
+ | |||
+ | |||
+ | :--->'''[[Team:Chiba/protocol/ligation/ligation|Ligation]]''' | ||
+ | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample No</td> | ||
+ | <td>(1)</td><td>(2)</td><td>(3)</td><td>(4)</td><td>(5)</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>insert①C0178</td> | ||
+ | <td>1</td><td>-</td><td>-</td><td>1</td><td>-</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>insert②C0170</td> | ||
+ | <td>-</td><td>1</td><td>-</td><td>-</td><td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>vector③J04500</td> | ||
+ | <td>-</td><td>-</td><td>2</td><td>2</td><td>2</td> | ||
+ | </tr>] | ||
+ | <td>ligase</td> | ||
+ | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Buffer</td> | ||
+ | <td>2</td><td>2</td><td>2</td><td>2</td><td>2</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>dH2O</td> | ||
+ | <td>6</td><td>6</td><td>5</td><td>4</td><td>4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td> | ||
+ | |||
+ | </tr> | ||
+ | </table> | ||
Revision as of 15:54, 3 September 2008
β班の実験ノート
Contents |
Week 1
- 17 August, 2008
- 18 August, 2008
- --->(20/8)Mini prep
- 19 August, 2008
- --->Gel Check
- 20 August, 2008
- (18/8)--->Mini prep
- --->Gel Check
Sample No | ① | ② | ③ | ④ |
Sample DNA | 3 | 3 | 3 | 3 |
Loading Dye | 2 | 2 | 2 | 2 |
dH2O | 7 | 7 | 7 | 7 |
TOTAL | 12 | 12 | 12 | 12 |
From the left; |
- 21 August, 2008
- (20/8)--->PCR
- --->Gel Check
From the left; |
- 22 August, 2008
- Transformation
- (Locate & 2 Punch ~iyama ver.~)
- 23 August, 2008
- --->Mini prep
- --->Digestion
- --->Gel Check</table>
From the left; |
|
Week 2
- 24 August, 2008
- (22/8)--->Mini prep
- --->Digestion
- --->Gel Check
Sample No | ① | ② | ③ | ④ | ⑤ | ⑥ | ⑦ | ⑧ |
Sample DNA | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Loading Dye | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
dH2O | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 |
TOTAL | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 |
From the left; (↓single digestuion) |
Sample No | ⑤ | ⑥ | ⑦ | ⑧ |
Sample DNA | 9 | 9 | 9 | 9 |
Loading Dye | 2 | 2 | 2 | 2 |
dH2O | 1 | 1 | 1 | 1 |
TOTAL | 12 | 12 | 12 | 12 |
From the left; (single digestuion) |
- 25 August, 2008
- (24/8)--->Digestion
Sample No | ① | ② | ③ | ④ |
Sample DNA | 5 | 5 | 1 | 1 |
EcoRⅠ | 0.5 | o.5 | - | - |
SpeⅠ | 0.5 | 0.5 | - | - |
Buffer 1 | 1 | 1 | - | - |
Buffer 3 | - | - | 1 | - |
BSA | 1 | 1 | - | - |
dH2O | 2 | 2 | 7.5 | - |
TOTAL | 10 | 10 | 10 | 1 |
- --->Gel Check
Sample No | ① | ② | ③ | ④ |
Sample DNA | 10 | 10 | 10 | 10 |
Loading Dye | 2 | 2 | 2 | 1 |
dH2O | - | - | - | 4 |
TOTAL | 12 | 12 | 12 | 6 |
From right; |
)--->Digestion
Sample No | ① | ② | ③ |
Sample DNA | 33ng/μl×60μl | 33ng/μl×60μl | 100ng/μl×20μl |
PstⅠ | 2 | 2 | 2 |
XbaⅠ | 2 | 2 | - | SpeⅠ | - | - | 2 |
Buffer 2 | - | - | 5 |
Buffer 3 | 10 | 10 | - |
BSA | 10 | 10 | 5 |
dH2O | 16 | 16 | 16 |
TOTAL | 100 | 100 | 52 |
- --->Gel Check
Sample No | ① | ② | ③ |
Sample DNA | 30 | 30 | 30 |
Loading Dye | 6 | 6 | 6 |
TOTAL | 36 | 36 | 36 |
|
|
|
- 26 August, 2008
(8/25)--->Gel Extract
- --->Zymo Clean
- ①5μl
- ②5μl
- ③27μl
- --->Gel Check
Sample No | ① | ② | ③ |
Sample DNA | 2 | 2 | 2 |
Loading Dye | 2 | 2 | 2 |
H2O | 8 | 8 | 8 |
TOTAL | 12 | 12 | 12 |
From left;
|
- --->Dephosphorylation
- ③
- --->Zymo Clean
- ③->9μl
- --->Gel Check
Sample DNA ③ | 1 |
Loading Dye | 1 |
dH2O | 4 |
TOTAL | 6 |
③ -> OK ・・・30ng? |
- --->Ligation
Sample No | (1) | (2) | (3) | (4) | (5) |
insert①C0178 | 1 | - | - | 1 | - |
insert②C0170 | - | 1 | - | - | 1 |
vector③J04500 | - | - | 2 | 2 | 2 | ligase | 1 | 1 | 1 | 1 | 1 |
Buffer | 2 | 2 | 2 | 2 | 2 |
dH2O | 6 | 6 | 5 | 4 | 4 |
TOTAL | 10 | 10 | 10 | 10 | 10 |
- 27 August, 2008
- PCR
- Colony PCR of 5 colonies from ligation plates (27/8) and one from control plate(BBa_F2620[158](2007)).
From left; |
From left; |
- 28 August, 2008]
From left; |
- 29 August, 2008
- 30 August, 2008
Week 3
- 31 August, 2008
- 1 September, 2008
- 2 September, 2008
- 3 September, 2008
- 4 September, 2008
- 5 September, 2008
- 6 September, 2008
Week 4
- 7 September, 2008
- 8 September, 2008
- 9 September, 2008
- 10 September, 2008
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