Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

Inoculation of colonies from plate with ligation of pSB2K3 + alpha_linker under PT7 (BBa_K103019) to LB with kanamycin.

Preparation of AraC+pBAD+AID

Michał K.

Inoculation of colonies from plate with ligation of pSB1A3 + AraC+pBAD+AID to LB with ampicillin.

Preparation of BioBricks

Michał K.

1. Inoculation of colonies from plate with ligation of 5kb(?????) + pT7_alpha to LB with kanamycin and RFP(?????) + AraC+pBAD+AID to LB with ampicillin.
2. Digest of 3kb(????)+pLac_OmpA_alpha and RFP(????)+OmpA with EcoRI and SacI (BamHI buffer).
3. Gel electrophoresis and gel-out of proper band 500 bp for RFP(????)+OmpA and 3000 bp for 3kb(????)+pLac_OmpA_alpha.
4. Ligation of DNA fragments: 3kb(?????)_ alpha + Omp.

Piotr

1. Transformation of TOP10 with above ligation.
2. Tranformants planting on LB with kanamycin.