Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.:

1. Isolation of hypothetical pMPM-T5 with transcription fusion.
2. Isolation of hypothetical pMPM-T5 with translation fusion.
3. Control digest of pMPM-T5 with transcription fusion with EcoRI (EcoRI buffer).
4. Control digest of pMPM-T5 with translation fusion with BamHI (BamHI buffer).
5. Choice of proper clones and preparation for sequencing.

Here we found the clone with translational fusion on pMPM. These are different isolations in each lane. We have been working with the AT6 ever since.
Digest was conducted with BamHI

2 to 6-Control digests of few clones with transcription fusion; 7-pMPMT5-AID; all plasmids digested with EcoRI.

Michał L., Ewa, Marcin

1. Inoculation of liquid LB with E.coli TOP10 carrying pMPM-T5+AID
2. Inoculation of liquid LB with E.coli TOP10 carrying pTrc99A+AID