Team:Warsaw/Calendar-Main/6 May 2008

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<h3>Preparation of <a href="https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID">pMPMT5+AID</a> construct</h3>
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<h4>Piotr, Weronika</h4>
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<p>1. Preparation of  electro- and chemocompetent bacteria <i>E. coli</i> TOP10 (lot of fun)<br>
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2. Isolation of genomic DNA (<i>E.coli</i> Rosetta)<br>
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<li>Preparation of <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrocompetent">electro-</a> and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemocompetent">chemocompetent</a> bacteria <i>E. coli</i> TOP10.</li>
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3. Setup of overnight cultures (pTrc99A-AID, pMPM-T5, pZC320)<br> </p>
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<li>Setup of overnight cultures (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a>, <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a>, <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a>) on LB broth with proper antibiotics.</li></ol><br>
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<h3> Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a> </h3>
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<h4>Michał L.</h4>
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#genomic_DNA_isolation"> Isolation of genomic DNA</a> (<i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>) - template DNA for PCR to obtain T7 polymerase DNA. DNA isolated from <i>E. coli</i> TOP10 strain was used as a negative control for PCR. <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_May_2008#fig1">Fig. 1</a>.<br>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/3/3c/Genomic_DNA_WAW.jpg"/></a>
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<var><b>Fig. 1.</b> Probes of isolated genomic DNA after mechanical shearing by pipetting. Without shearing the DNA wouldn't be visible in this kind of gel because genomic DNA is too big. 1 and 3-<i>E.coli</i> Rosetta DNA; 2-DNA ladder; 4-<i>E.coli</i> TOP10 DNA.</var>
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Latest revision as of 13:42, 26 October 2008

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Preparation of pMPMT5+AID construct

Piotr, Weronika

  1. Preparation of electro- and chemocompetent bacteria E. coli TOP10.
  2. Setup of overnight cultures (pTrc99A-AID, pMPM-T5, pZC320) on LB broth with proper antibiotics.

Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał L.

Isolation of genomic DNA (E.coli Rosetta and TOP10) - template DNA for PCR to obtain T7 polymerase DNA. DNA isolated from E. coli TOP10 strain was used as a negative control for PCR. Fig. 1.

Fig. 1. Probes of isolated genomic DNA after mechanical shearing by pipetting. Without shearing the DNA wouldn't be visible in this kind of gel because genomic DNA is too big. 1 and 3-E.coli Rosetta DNA; 2-DNA ladder; 4-E.coli TOP10 DNA.

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