Team:Lethbridge/Notebook/Lab Work/Group3

From 2011.igem.org

(Difference between revisions)
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=June= <br />
=June= <br />
-
==June 21st== <br />
+
==June 21st, 2011== <br />
Restriction, Ligation, and Transformation <br />
Restriction, Ligation, and Transformation <br />
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#* Restricted destination plasmid pSB1C3 with EcoR1 and Pst1 <br />
#* Restricted destination plasmid pSB1C3 with EcoR1 and Pst1 <br />
-
## [[Team: Lethbridge/Notebook|Ligation]]
+
## [[Team: Lethbridge/Notebook|Ligation]]<br />
#* Ligated lumazine synthase-dt into destination plasmid pSB1C3
#* Ligated lumazine synthase-dt into destination plasmid pSB1C3
#* Ligated pBAD-RBS into destination plasmid pSB1C3
#* Ligated pBAD-RBS into destination plasmid pSB1C3
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### [Team:Lethbridge/Notebook|Transformation]]
### [Team:Lethbridge/Notebook|Transformation]]
#* Transformed lumazine synthase - dt in pSB1C3 into DH5&prop;
#* Transformed lumazine synthase - dt in pSB1C3 into DH5&prop;
 +
#* Transformed pBAD-RBS in pSB1C3 into DH5&prop;
 +
 +
==June 24th, 2011==
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 +
Picked cells from June 21st transformation <br / >
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 +
==June 25th, 2011==
 +
 +
Minipreped Cells picked on June 25th, 2011 < br />
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 +
==June 28th==
 +
 +
Restriction digest and ran on gel
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Lumazine - dt construct successfully inserted into pSB1C3 <br />
 +
Glycerol stocked.
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=ADD GEL PICTURE=
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==July 8th==
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Performed antibiotic assay on lumazine - dt in pSB1C3 <br />
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Samples from one colony grew in Kanamicyn; this stock was discarded
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 +
PCR: PCR'd out RFP and lumazine

Revision as of 02:00, 15 September 2011



=June=
==June 21st, 2011==

Restriction, Ligation, and Transformation
Lumazine synthase - double terminator and pBAD - RBS into pSB1C3

  1. Restriction
    • Restricted lumazine synthase - dt out of pSB1K3 and pBAD - RBS out of pSB1T3 with EcoR1 and Pst1
    • Restricted destination plasmid pSB1C3 with EcoR1 and Pst1
    1. Ligation
    • Ligated lumazine synthase-dt into destination plasmid pSB1C3
    • Ligated pBAD-RBS into destination plasmid pSB1C3
      1. [Team:Lethbridge/Notebook|Transformation]]
    • Transformed lumazine synthase - dt in pSB1C3 into DH5∝
    • Transformed pBAD-RBS in pSB1C3 into DH5∝

Contents

June 24th, 2011

Picked cells from June 21st transformation

June 25th, 2011

Minipreped Cells picked on June 25th, 2011 < br />

June 28th

Restriction digest and ran on gel Lumazine - dt construct successfully inserted into pSB1C3
Glycerol stocked.

ADD GEL PICTURE

July 8th

Performed antibiotic assay on lumazine - dt in pSB1C3
Samples from one colony grew in Kanamicyn; this stock was discarded

PCR: PCR'd out RFP and lumazine
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