Team:Paris/August 21

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(→Screening of the cloning of E0240 and FlhDC+promotor)
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|100 bp DNA ladder
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Revision as of 14:05, 21 August 2008

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Contents

Construction of pFlgA - YFP tripart (+/- LVA)

Aim : Construction of "pFlgA-RBS-YFP-dbl ter" (pFlgA-E0430/E0432) Part icon regulatory.pngPart icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png

Digestion

Gel Extraction

Protocol [[Image:KR00019b.jpg| thumb|Gel Extraction of D166-D167]

Well 1 2 3 4 5 6 7 8 9 10 11 12
Sample 1kb ladder MP165.1 MP166.1 no sample D166 no sample D167 no sample 100pb ladder no sample
Expected size (pb) 2 957 2 996 2942 2981
Measured size (pb) 3 000 3 000

Screening of the cloning of E0240 and FlhDC+promotor

We obtained single colonies with the dilution 100.
13 clones were analysed by PCR

PCR screening

reaction mixture (25 µL)

  • 12,5 µL Quick load PCR Mixture 2X
  • 0,5 µL O18
  • 0,5 µL O19
  • 11,5 µL water

PCR screening programm

  • elongation time: 1 min 30
  • primers: O18 and O19
Gel n°1 (E0240 in pSB3K3)
well n° 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
sample 1 kb DNA ladder positive control negative control S159.1 100 bp DNA ladder
colonie n° 1 2 3 4 5 6 7 8 9 10 11 12 13
expected size
measured size
Gel n°2 (FlhDC+promotor in pSB1A2)
well n° 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
sample 1 kb DNA ladder positive control negative control S161.1 100 bp DNA ladder
colonie n° 1 2 3 4 5 6 7 8 9 10 11 12 13
expected size
measured size
Gel n°1 (E0240 screening)
Gel n°2 (FlhDC+promotor screening)
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