Lan:LE daily entries

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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

  • 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)



  • 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

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