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Team:NTU-Singapore/Notebook/24 June 2008
From 2008.igem.org
- Chin Chong
- Plates with BL21 cells and Top10 cells both with LacI-GFP appears pinkish in normal light after overnight incubation
- Under UV the redness becomes more obvious
- Strange observation as it supposedly is Green Fluorescene and not Red Fluprescene
- Received quotations for Lysis gene and LsrA gene synthesis from 1st Base, AtiBioTech
- Redesigned primers of E7&Imm
- Conduct Trial for Characterization of LacI-GFP with M9 medium
- Use the M9 salts that were prepared the day before
- Add glycerol as Carbon source
- Inoculate BL21 and Top10 cells into two seperate 50ml tubes and incubate at 37 deg cel
- Measure the OD at 600nm from time zero=time of incubation at hourly intervals
- Results showed that the growth rate of cells in M9 medium is slow
- Inoculate Top10 cells with LacI-GFP in 5ml of LBA, 5ml of M9 with LBA and 5ml of M9 in three different 50 ml tubes
- Inoculate BL21 cells with LacI-GFP in 5ml of LBA
- Prepared 20 LBA agar plates for future use
- Plates with BL21 cells and Top10 cells both with LacI-GFP appears pinkish in normal light after overnight incubation
- Hung (4pm-530pm): transformation of 7 plasmids from the Registry:
- LacI-GFP [http://partsregistry.org/wiki/index.php?title=Part:BBa_I763004 BBa_I763004]
- GFP producer [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840 BBa_E0840]
- GFP reporter device [http://partsregistry.org/wiki/index.php/Part:BBa_E0240 BBa_E0240]
- BBa_I20260: [http://partsregistry.org/Part:BBa_J23101 BBa_J23101 standard promoter]-[http://partsregistry.org/wiki/index.php/Part:BBa_E0240 BBa_E0240 GFP reporter device]
- [http://partsregistry.org/Part:BBa_I20269 BBa_I20269],[http://partsregistry.org/Part:BBa_I20270 BBa_I20270],[http://partsregistry.org/Part:BBa_I20268 BBa_I20268]: GFP reporter devices with Weak (BBa_J23150), medium (BBa_J23151) and strong (BBa_J23102) promoters respectively.
