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Team:NTU-Singapore/Notebook/25 June 2008
From 2008.igem.org
Wednesday 25 June
- 1030am: Hung: take out the agar plates incubated from Tuesday night. For details of the plasmids, please refer to Tuesday notebook
- GFP reporters with standard, weak, medium and strong promoters all have colonies.
- GFP reporter device BBa_E2040 however shows no colony.
- GFP producer BBa_E4080 shows a few colonies.
- LacI-GFP shows no colony.
- GFP self-ligation shows a few colonies.
- pFe-GFP shows no colony.
- 11am: Group meeting to discuss about current obstacles encountered during wetlab.
- 12:15pm: Choon Kit: digestion for:
- pFe with SpeI/PstI
- GFP with XbaI/PstI
- pFe with SpeI to check SpeI activity
- Terminator with SpeI to check SpeI activity
Incubate for 3 hours.
- 3:30pm: Hung, Min-Lin: Transform these following plasmids to Homemade top10 cells for amplification and obtaining new plasmids with higher purity (using new Miniprep Kit):
- pT7 (ampR)
- LacI (ampR)
- RBS 34 (ampR)
- RBS 32 (ampR)
- GFP E0240 (ampR)
- Terminator (AK resistance)
- pFe (ampR)
- empty plasmid with AmpR
- 6pm: Hung: cell cloning on agar plates for the above plasmids
- 6:30pm: Hung: inoculate in 5ul LBA each for:
- LacI-GFP in top10
- LacI-GFP in BL21 cell
- self-ligated GFP cell
- E0480 GFP producer cell
- Choon Kit: 3pm to 7pm:
- Gel electrophoresis: controls of pFe and Terminator each show only 1 single band, which mean there's nothing wrong with SpeI.
- Gel extraction of pFe (cut with SpeI and PstI) and GFP (cut with XbaI and PstI).
- Gel electrophoresis again to check the extracted plasmids.
