Team:NTU-Singapore/Notebook/12 September 2008

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m (New page: =Friday 12 September:= *1200am: punching and dilution into TE buffer at 50 degrees C of [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=pSB2K3 pSB2K3] (for obtaining plasmid vector...)
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=Friday 12 September:=
=Friday 12 September:=
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*1200am: punching and dilution into TE buffer at 50 degrees C of [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=pSB2K3 pSB2K3] (for obtaining plasmid vector with KanR).
*1200am: punching and dilution into TE buffer at 50 degrees C of [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=pSB2K3 pSB2K3] (for obtaining plasmid vector with KanR).
*1230pm: digestion with E/P for Detection system (LsrA-RBS-Lysis-Term) (insert) and pSB2K3 (vector). Incubate for 2 hours until 3pm.
*1230pm: digestion with E/P for Detection system (LsrA-RBS-Lysis-Term) (insert) and pSB2K3 (vector). Incubate for 2 hours until 3pm.
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**LsrA-RBS-Lysis-Term (insert) ligated with pSB2K3 (vector) into top10 (and use LBK plate for selection).
**LsrA-RBS-Lysis-Term (insert) ligated with pSB2K3 (vector) into top10 (and use LBK plate for selection).
**Detection system (LsrA-RBS-Lysis-Term) and Production system (LacI-RBS-E7-Term) into LuxS k/o (LBA plates).
**Detection system (LsrA-RBS-Lysis-Term) and Production system (LacI-RBS-E7-Term) into LuxS k/o (LBA plates).
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Revision as of 13:09, 25 October 2008

Friday 12 September:

  • 1200am: punching and dilution into TE buffer at 50 degrees C of [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=pSB2K3 pSB2K3] (for obtaining plasmid vector with KanR).
  • 1230pm: digestion with E/P for Detection system (LsrA-RBS-Lysis-Term) (insert) and pSB2K3 (vector). Incubate for 2 hours until 3pm.
  • 2pm-530pm: prepare LBK plates.
  • 3pm-415pm: MinElute purification and Ligation (followed by another purification) for the insert and vector digested at 1230pm.
  • 4pm: punching for [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0430 BBa_E0430] (RBS-YFP-Term) and [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=BBa%20E0420 BBa_E0420] (RBS-CFP-Term)
  • 430-6pm: Transformation for:
    • RBS-YFP-Term and RBS-YFP-Term into Top10 cells. (LBA plates)
    • LsrA-RBS-Lysis-Term (insert) ligated with pSB2K3 (vector) into top10 (and use LBK plate for selection).
    • Detection system (LsrA-RBS-Lysis-Term) and Production system (LacI-RBS-E7-Term) into LuxS k/o (LBA plates).
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