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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)

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

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