Team:Warsaw/Calendar-Main/27 September 2008

Mutagenesis of protein A Paweł

Isolation of plasmid DNA from overnight cultures (it was really hard work...). Control digest of isolated plasmids with KpnI and BamHI (if mutagenesis really occured, ~550bp band should be visible). Results of digest:  ADelL+KpnI and ADelP primers mutagenesis:  pACYClac+ompA-&Delta;A-omega: 0 positive, 5 negative pACYClac+ompa-&Delta;A-alpha</a>: 3 positive, 7 negative</li> pACYClac+OmpA-omega-&Delta;A</a>: 1 positive, 2 negative</li> </ul> AMutL+KpnI and AMutP mutagenesis:</li> <ul> pACYClac+ompA-&Delta;A-omega</a>: 4 positive, 16 negative</li> pACYClac+ompa-&Delta;A-alpha</a>: 4 positive, 8 negative</li> pACYClac+OmpA-omega-&Delta;A</a>: 2 positive, 15 negative</li> </ul> </ul> Plasmids which gave positive digest results prepared for sequencing (one sample from each variant which gave positive result).</li> </ol>