Lab Notes
[Abbreviation notes: 4L = part BBa_K325909 (LuxCDABEG), 2D = part BBa_K325219 (Red firefly luciferase and LRE), 4N = part BBa_K325100 (EPIC firefly luciferase and LRE); Concentration of plasmids are in units of ng/μL (if not mentioned)]
July
7/17
- I7 test tubes, 4 monoclone (4L, 2DH, 2DH-5)
- 2D and 4N transformation (2DH,2DH-5)
7/18
- Plasmid extraction for 7 test tubes
- PCR 4L and LuxG (failed)
7/19
- Plasmid extraction
7/21
- Four Agrobacteria transformations for 2*4L, 1*2D and 1*4N
7/25
- Transformed 2*LuxG, 2*4L, 2*2D, 2*4N, totaling up to 9 petri dishes
- 2*LuxG 2*4L 1*2D 1*4N totaling up to 6 test tubes
7/26
- PCR LuxG with primers
- Added 4L monoclone into 2 Erlenmeyer flasks
- Used XBaI and PstI to digest the 4L monoclone
- Extracted plasmids (1*4N, 1*2D, 2*4L, 2*LuxG)
- Gel electrophoresis the products from PCR and enzyme digestion.
7/27
- LuxG gel extraction (digested by XBaI and PstI)
- Tested for concentration
7/28
- Took 20ml of 4L from the Erlenmeyer flask
- Extracted plasmid (4*2D, 3*4L)
- Conducted gel electrophoresis to confirm plasmid size
- Tested for concentration
- 4L succeeded, but the concentration of 2D was too low for gel electrophoresis
- Added 8mL of 0.1M Arabinose into a 80mL flask and 10ml of 0.1 Arabinose into a100mL flask
- After 3.5 hours, visible light was emitted
- Gradient test of 4L with 0.1M Arabinose
7/29
- Prepared 3L of LB, but had to be redone due to an inaccurate electronic balance
- Prepared 2L liquid LB, 1 L solid LB. All sterilized.
- Washed and sterilized 62 test tubes.
- Organized plasmids and DNA
- Designed further experiments with guidance professors.
August
8/1
- Extracted 8 tubes of pHB plasmid
- Concentration: 67.439, 72.8, unknown, 128.6, 203.2, 153.6, 101.2, 61.7
- Digested pHB using BamHI and PstI, ran gel electrophoresis and did gel extraction
- Digested 4L using BamHI and PstI, ran gel electrophoresis and did gel extraction
- Monoclone 4L into 2 flasks and 4 test tubes
- Gradient test
- 0.1M solution emitted the brightest light.