Team:Paris/27 August 2009

From 2009.igem.org

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August 27th

Lab work

PCR on OmpA signal/g3p colony

Total volume = 25ul

2XSuper mix : 12,5 ul

Diluted bacteria : 5 ul

VF2 (10 uM) : 0,5 uL

VR (10uL) : 0,5 uL

H2O : 6,5 uL



gel purification

gel purif using promega kit



PCR


Pcr of:

  • FecR part A & B
  • Tg3p


purification on column


Purif of:

  • OmpA linker (SacI/P) Failed (wrong size and two band)
  • FecI (X/P) Failed (no DNA ?! )
  • pBad (S/P) Failed (no DNA ?! )


Transfo

Jun Fos AIDA gen

Transfo

A40 TolRII: topo vector

O/N

A41 OmpAsignal : topovector