Team:Warsaw/Calendar-Main/18 July 2008

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<h3>Cloning of protein A DNA to OmpA constructs</h3>
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<li>Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
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<p>'''Cloning of protein A DNA to OmpA constructs'''<br>
 
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1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
 
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a>
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+SacI">AL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a>
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2. Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin. <br>
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<li> Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin. </li></ol>
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<p>Paweł: pET15b vector purified.
<p>Paweł: pET15b vector purified.
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Digested of pET15b and pACYClac+OmpA+omega with BamHI and NdeI<br>Gel-out of OmpA+omega (cutted from pACYClac+OmpA+omega, ~1350 bp)</p>
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Digest of pET15b and pACYClac+OmpA+omega with BamHI and NdeI<br>Gel-out of OmpA+omega (cutted from pACYClac+OmpA+omega, ~1350 bp)</p>
<p>Ligation of gel-outed fragment into digested pEt15b
<p>Ligation of gel-outed fragment into digested pEt15b
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Revision as of 18:21, 1 October 2008

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Cloning of protein A DNA to OmpA constructs

  1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used: AL+SacI and AP+NotI
  2. Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin.

Paweł: pET15b vector purified. Digest of pET15b and pACYClac+OmpA+omega with BamHI and NdeI
Gel-out of OmpA+omega (cutted from pACYClac+OmpA+omega, ~1350 bp)

Ligation of gel-outed fragment into digested pEt15b