Team:NTU-Singapore/Notebook/25 June 2008

From 2008.igem.org

(Difference between revisions)
m
Line 15: Line 15:
**pFe with SpeI to check SpeI activity
**pFe with SpeI to check SpeI activity
**Terminator with SpeI to check SpeI activity<br>Incubate for 3 hours.
**Terminator with SpeI to check SpeI activity<br>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.

Revision as of 13:03, 25 June 2008

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.
Retrieved from "http://2008.igem.org/Team:NTU-Singapore/Notebook/25_June_2008"