Lan:LE daily entries

From OpenWetWare
Jump to navigationJump to search

Lan yellow8x3.png

Home        Who We Are        Research        Publications        Lab Members        Protocols and Recipes        Lab Members Only       


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)

Results

AP06-52.jpg

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

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