Team:Warsaw/Calendar-Main/8 October 2008

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<h3>Preparation of vector for pT7 constructs</h3>
<h3>Preparation of vector for pT7 constructs</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<p>Inoculation of colonies from plate with ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI to liquid LB + ampicillin. </p>
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<p>Inoculation of colonies from plate with ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> (with removed XbaI site) to liquid LB + ampicillin. </p>

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Preparation of linker_alpha (BBa_K103009)

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.

Preparation of linker_omega (BBa_K103013)

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
Fig. 23.traw_10_10_2008.jpg

Preparation of OmpA-linker-omega-linker (BBa_K103016)

Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA-linker-omega-linker (BBa_K103016)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.

Preparation of vector for pT7 constructs

Michał K.

Inoculation of colonies from plate with ligation of pET15b+OmpA_omega (with removed XbaI site) to liquid LB + ampicillin.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Piotr

Inoculation of colonies from plate with ligation of pSB2K3 + BBa_K103018 (without internal EcoRI site) to liquid LB + kanamycin.

Preparation of AID(BBa_K103001)

Michał K.

  1. Colony PCR with AIDlNrH and AIDpLinB primers on colonies from plates with transformations pSB1A3+AID(BBa_K103001) (annealing temperature - 55°C, 60 s of elongation step).
  2. Inoculation of confirmed colonies to liquid LB + ampicillin.
Fig. 14.Kolonijny_aid_07_10_2008.jpg

Preparation of AID under pBAD/araC (BBa_K103002)

Piotr

  1. Transformation of TOP10 with ligation pMPMT5+AID (with removed EcoRI site).
  2. Tranformants plating on LB with tetracycline.