Lan:LE daily entries: Difference between revisions
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**State what are your finding and any comments | **State what are your finding and any comments | ||
<h1>2008/ | <h1>2008/07/01 - '''AP06-52'''</h1> | ||
<h2>Expression assay on positives clones</h2> | <h2>Expression assay on positives clones</h2> | ||
Digestion positives clone('''ND01-XX and ND01-YY''') where tested for protein expression | Digestion positives clone('''ND01-XX and ND01-YY''') where tested for protein expression | ||
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<td> 8µL </td> | <td> 8µL </td> | ||
</tr> | </tr> | ||
Notes: | |||
*Samples in lanes 2 and 6 had very low biomass. Apparently, #6 was for miniprep and not for protein expression. | |||
*Sample #12 was grown in SOC media (w/o glucose and pH was 8.75) | |||
<h2> Minipreps on recalcitrants clones!!</h2> | |||
*minipreps were done on clones B4-S, B6-s and B14-L | |||
Notes: Miniprep should have been done on B12-L and not B6-S |
Latest revision as of 06:40, 2 July 2008
Please follow this format for your entries
- Date(year/month/day)-Hard copy lab book reference (Initials lab book #-page)
- Title of experiment with brief description
- Experimental procedures: Refer to appropriate protocol and note any changes in them
- Results: Post pictures with caption and appropriate identification of lanes on gels
- State what are your finding and any comments
2008/07/01 - AP06-52
Expression assay on positives clones
Digestion positives clone(ND01-XX and ND01-YY) where tested for protein expression
Experimental procedures
- Used O/N culture for miniprep to inoculate 10 mL 2XYTGB flasks(125 mL) to OD~0.4
- Induced with 0.1 mM IPTG (100 µL of 100 mM stock)
- Incubated in shaker B at 37°C O/N
- Harvested at 7:30 and lysed using lysis buffer (10X stock diluted to 1X, nothing else)
Results
Notes:- Samples in lanes 2 and 6 had very low biomass. Apparently, #6 was for miniprep and not for protein expression.
- Sample #12 was grown in SOC media (w/o glucose and pH was 8.75)
Minipreps on recalcitrants clones!!
- minipreps were done on clones B4-S, B6-s and B14-L
Notes: Miniprep should have been done on B12-L and not B6-S
Lane | Sample | Volume |
---|---|---|
1 | A4-S PS | 10 µL |
2 | A9-S | 10 µL |
3 | A21-L | 10 µL |
4 | A22-L | 10 µL |
5 | B3-S | 10 µL |
6 | B12-L | 10 µL |
7 | B16-L | 10 µL |
8 | C32-S | 10 µL |
9 | C33-S | 10 µL |
10 | C34-S | 10 µL |
11 | C36-S | 10 µL |
12 | C7-S | 10 µL |
13 | C42-L | 10 µL |
14 | C44-L | 10 µL |
15 | PS MARKER | 8µL |