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

Piotr

Design of primers for sequencing something we want from pMPM-T5.
Primers:

• pMPMT5seqF
• pMPMT5seqR
• AIDseqP

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. Gel electrophoresis (no proper colonies found). Fig. 1.
Fig. 1. 1-DNA ladder;
2 and 3-hypothetical pMPM-T5 with transcription fusion digested with EcoRI (2 clones);
4-pMPM-T5-AID digested with EcoRI;
5-hypothetical pMPM with translation fusion digested with BamHI;
6-pMPM-T5-AID digested with BamHI.