Team:Warsaw/Calendar-Main/14 October 2008

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Preparation of linker_alpha (BBa_K103009)

Piotr

  1. Transformation of TOP10 with overnight ligation - pSB2K3 + linker_alpha (BBa_K103009).
  2. Tranformants plating on LB with kanamycin.

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

Michał K.

  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 - still no proper clones found. Fig. 1.
Fig. 1.EcoRI/PstI digests of isolated plasmids.
1. Marker
2-8.EcoRI/PstI digests of pACYC177+OmpA-linker-omega-linker plasmids

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

Isolation of plasmid from culture inoculated on previous day - pSB1A3 carrying alpha_linker under PT7 (BBa_K103019).

Preparation of omega_linker under PT7 (BBa_K103020)

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + omega_linker under PT7 (BBa_K103020)).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. Fig. 2.
Fig. 2.EcoRI/PstI digests of isolated pSB2K3+omega_linker under PT7 (BBa_K103020) plasmids
1. Marker
2-4. Digested plasmids pSB2K3+omega_linker under PT7 (BBa_K103020)

Preparation of AID under pBAD/araC (BBa_K103002)

Michał K.

  1. Transformation of TOP10 with overnight ligation - pSB1A3 + AID under pBAD/araC (BBa_K103002).
  2. Tranformants plating on LB with ampicillin.

Preparation of OmpA_linker_alpha_linker under Plac (BBa_K103017)

Piotr

  1. Transformation of TOP10 with overnight ligation - pSB2K3 (with OmpA_linker and lactose promoter) + alpha2.
  2. Tranformants plating on LB with kanamycin.

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