Team:NTU-Singapore/Notebook/24 June 2008



  

=Tuesday 24 June=

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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 BBa_I763004
 * GFP producer BBa_E0840
 * GFP reporter device BBa_E0240
 * BBa_I20260: BBa_J23101 standard promoter-BBa_E0240 GFP reporter device
 * BBa_I20269,BBa_I20270,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.