Team:Warsaw/Calendar-Main/2 September 2008

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A-alpha, Z-alpha and Z-omega from overnight culture inoculated in fresh LB and cultured until OD=0,5
Cultures induced with different concentrations of IPTG in 22C and 37C.
Samples were collected twice: after 3h and next day (samples were centrifuged and frozen)

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 <li>Samples were collected twice: after 3h and next day (samples were centrifuged and frozen)</li></ol></p>
-<h3>Mutagenesis of protein A<br>Paweł</h3>
-<p>Mutagenesis of protein A was performed using 2 pairs of primers: <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/primers#ADelL+KpnI>ADelL+KpnI</a> and <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/primers#ADelP>ADelP</a> (deletion of aminoacids involved in interaction with protein Z) and <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/primers#AmutL+KpnI>AMutL+KpnI</a>  and <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/primers#AmutP>AMutP</a> (changing of few aminoacids involved in interaction with protein Z).</p>
-<p>Mutagenesis were performed on 3 vectors:  pACYClac+ompA-A-omega, pACYClac+ompa-A-alpha and pACYClac+ ompa-alpha-A.</p>
-<p>Each mutagenesis were performed using each primer at final concentration 0,1 uM, dNTPs at final concentration 0,25 uM and Walk (Pwo) polymerase (shipped by <a href=http://aabiot.com/>A&A Biotechnology</a>), with 100 ng of DNA template. </p>
-<p><br>PCR program (15 cycles):
-<pre style="text-align: left">94C 5 min
-C 30 s
-C 30 s
-C 10 min
-C 8 min
-C pause
-</pre></p>
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