Lan:LE daily entries
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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
2008/06/30 - AP06-39
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
- 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 |
