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

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.

Choon Kit

• • Digestion of pFe with SpeI/PstI, with SpeI (control) and with PstI (control).
  • Digestion of GFP producer with XbaI/PstI and with EcoRI/PstI(control).
  • Gel electrophoresis: sample cut with SpeI alone got smeared though only digested for 2 hours.
  • Gel extraction.
  • Ligation of pFe and GFP producer, cell ligation of GFP as a control.
  • Cell cloning of pFe-GFP, GFP self-ligation and 7 plasmids transformed by Hung.