DNA (PCR products from previous day) gel electrophoresis and isolation of DNA from proper bands.
Digestion of DNA with HindIII and SalI.
Digestion of pMPM-T5 + AID with HindIII and SalI.
Clean-up of products of p.2 and p.3 reactions.
Ligation of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion).
Transformation of E.coli TOP10 with ligation product.
Plating transformants on LB+tetracycline.
Digestion of pMPM-T5 + transcription fusion with EcoRHI and HindIII.
Use of T4 polymerase for blunting.
DNA gel electrophoresis.
Isolation of DNA from proper band.
Ligation of product p.11 (pMPM-T5 + T7 RNA-polymerase).
Transformation of E.coli TOP10 with ligation product.
Plating transformants on LB + tetracycline.