Team:Groningen/Labwork/21 August 2013

Sander

made 8 inoculations of j04450 strain in LB-Broth. made inoculations of CheY knockout and wild type bacillus on 0.3% and 0.4% agar with LB nutrient.

Inne

Inoculated the 0.3% and 0.4% agar plates with wt and delta cheY colonies from the plates from 19-08 (same as 0.7% plates). Each strain was grown in duplo. Plates were left to grow overnight in the 37C incubator.

Claudio

The synthetic gene sequences, which were ordered, are 16 and I'll refer to that as S# (see LINK).

S1, S2, S5, S6 and pSB1C3 were digested (NEB enzymes) with EcoRI and PstI.
S16 was digested (FastDigest) with EcoRI and BamHI.
S3 and S4 were digested (FastDigest) with BamHI and PstI.

S1, S2, S5, S6, SS1 (S16 + S3) and SS2 (S16 + S4) were ligated into pSB1C3.
The ligation reactions were transformed into E. Coli DH5α and plated on LB + Cm.