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Team:Warsaw/Calendar-Main/3 June 2008
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Attribution analyzer
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Interactive list of topics
Preparation of T7 constructs
Michał K.:
DNA (PCR products from previous day) gel electrophoresis and
isolation
of DNA from proper band.
Digest
of DNA with HindIII and SalI (2x Tango buffer).
Digest
of pMPM-T5 + AID with HindIII and SalI.
Clean-up
of reaction products.
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.
Digest
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.