Team:NTU-Singapore/Notebook/24 June 2008

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*Chin Chong
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=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
**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
***Under UV the redness becomes more obvious
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**Inoculate BL21 cells with LacI-GFP in 5ml of LBA
**Inoculate BL21 cells with LacI-GFP in 5ml of LBA
**Prepared 20 LBA agar plates for future use
**Prepared 20 LBA agar plates for future use
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*Hung (4pm-530pm): transformation of 7 plasmids from the Registry:
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===Hung===
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(4pm-530pm): transformation of 7 plasmids from the Registry:
**LacI-GFP [http://partsregistry.org/wiki/index.php?title=Part:BBa_I763004 BBa_I763004]
**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 producer [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840 BBa_E0840]
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**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]
**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.
**[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.
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*Choon Kit:
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===Choon Kit===
**Digestion of pFe with SpeI/PstI, with SpeI (control) and with PstI (control).
**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).
**Digestion of GFP producer with XbaI/PstI and with EcoRI/PstI(control).
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**Ligation of pFe and GFP producer, cell ligation of GFP as a control.
**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.
**Cell cloning of pFe-GFP, GFP self-ligation and 7 plasmids transformed by Hung.
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Revision as of 11:28, 25 October 2008

Contents

Tuesday 24 June

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.
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