Team:Warsaw/Calendar-Main/3 June 2008

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DNA (PCR products) 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