Preparation of linker_alpha (BBa_K103009)

Michał K.

1. Isolation of plasmids from culture inoculated on previous day.
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. Fig. 1.

Fig. 1.EcoRI/PstI digests of isolated plasmids
1. Marker
2-7. Digests of pSB1A3 carrying alpha_linker under PT7 (BBa_K103019)
8-11. Digests of pSB2K3 carrying alpha_linker under pLac
11-17. Digests of pSB2K3 carrying linker_alpha

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

Michał K.

Inoculation of colonies from plate with new ligation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to LB with kanamycin.

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

1. Control digest of isolated plasmid - pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) fragment (14 October) with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found. Fig. 1.
2. Overnight digest pSB1A3 carrying alpha_linker under PT7 (BBa_K103019) with EcoRI and BcuI (BamHI buffer).

Fig. 1.EcoRI/PstI digests of isolated plasmids
1. Marker
2-7. Digests of pSB1A3 carrying alpha_linker under PT7 (BBa_K103019)
8-11. Digests of pSB2K3 carrying alpha_linker under pLac
11-17. Digests of pSB2K3 carrying linker_alpha

Preparation of AID under pBAD/araC (BBa_K103002)

Michał K.

1. Isolation of plasmids from culture inoculated on previous day (pSB1A3 + AID under pBAD/araC (BBa_K103002)).
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found. Fig. 2.
3. Repetition of overnight ligation of isolated DNA fragments: pSB1A3 + AID under pBAD/araC (BBa_K103002).

Preparation of OmpA_linker_alpha_linker under Plac (BBa_K103017)

Michał K.

1. Isolation of plasmids from culture inoculated on previous day - pSB2K3 + OmpA_linker_alpha_linker under Plac (BBa_K103017)
2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - we found good clones. Fig. 1.