Team:Gifu/Notebook

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August

August 11th

Restriction Enzyme Digestion

samplerestriction enzymes
His-RFP with stop codonSpeI,EcoRI
5'-intron MSpeI,EcoRI
Lac-3'intron-RBSSpeI,EcoRI
His-RFP without stop codonXbaI,EcoRI
RBS-AmpSpeI,EcoRI

Ligation

  • EcoRI-His-RFP(with stop codon)-SpeI + XbaI-DT-Cm-EcoRI
  • EcoRI-5'intron M-SpeI + XbaI-DT-Cm-EcoRI
  • EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(without stop codon)-5'intron-DT-Cm-EcoRI
  • EcoRI-His-RFP(without stop codon)-SpeI + SpeI-Amp-EcoRI

Transformation

  • His-RFP(with stop codon)-DT-Cm
  • 5'intron M-DT-Cm
  • Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm
  • His-RFP(without stop codon)-Amp

Densitometry

sampleconcentration(ng/µL)
His-RFP(with stop codon) EcoRI-SpeI162.6
5'intron M EcoRI-SpeI45.4
Lac-3'intron-RBS EcoRI-SpeI120.9
His-RFP(without stop codon) EcoRI-SpeI160.0
His-RFP(without stop codon)-5'intron-DT-Cm XbaI-EcoRI 174.0
Amp EcoRI-SpeI101.8

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August 12th

Colony PCR

Electrophoresis

Shaking culture

To analysis sequence

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August 13th

Miniprep

PCR

Electrophoresis

Restriction Enzyme Digestion

Ligation

Transformation

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August 16th

Liquid Culture

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August 17th

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August 18th

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August 19th

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August 20th

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