1. The electroporations of 26/08 were checked.
   • had some colonies. they were ented in liquid culture.
   • LigC ( + ) did not grow. we figured that an insert of 35bp was too short to ligate so we decided to use as insert and + as vector . The following restriction digest was started:
     • was cut with SpeI and PstI
     • was cut with XbaI and PstI
2. The miniprep that was made to sequence the LigA construct on 26/08 was used again to perform a restriction digest (EcoRI and PstI). This was put on gel to check whether there actually was LigA-insert in the vector and whether the insert had the right length. The gel showed a signal at 1000 bp and at 2000 bp which coincides with the insert (BLp + GFP) and the vector.
3. A new setup to light the E.coli was engineered.
   • Fresh cultures were made from the old ones (LB plate ligA 14/08 and the two liquid cultures from 26/08 were used as templates.)
   • They were put in the 16°C room for about an hour
   • A blue light (40W) was put on them for about an hour
   • They were put in the 37°C incubator overnight

• Restriction4Real: we performed a gel electrophoresis together with opened plasmids (cut with EcoRI) to compare sizes

• Gel signals were inconclusive: gel ran not long enough

• Because of bad results yesterday the transformation of W was extra checked by performing a new PCR with different primers :

sample A : M13forward - virAforward sample B : M13forward - virAreverse sample C : virAforward - virAreverse sample D : M13reverse - virAforward sample E : M13reverse - virAreverse

Afterwards was gelelectrophoresis done and this showed virA was not present in the cell.

• ligation of X and Y showed a pover result so new ligations were done for virB and virG