Lan:LE daily entries

 Lab note books 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 Results
 * 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)

SDS-PAGE 15% 60 min

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

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