Team:Goettingen/week5-3
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<b>Insertion of <i>tar</i> into pUC18</b><br> | <b>Insertion of <i>tar</i> into pUC18</b><br> | ||
<ul> | <ul> | ||
- | <li>Experiment: <br>Double digest of <i>tar</i> and pUC18 using <i>Eco</i>RI, <i>Pst</i>I and Orange buffer (ThermoScientific) according to <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>. Gel extraction and purification of pUC18 and <i>tar</i> using peqGOLD | + | <li>Experiment: <br>Double digest of <i>tar</i> and pUC18 using <i>Eco</i>RI, <i>Pst</i>I and Orange buffer (ThermoScientific) according to <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>. Gel extraction and purification of pUC18 and <i>tar</i> using peqGOLD Gel Extraction Kit (Peqlab). Ligation of <i>tar</i> into pUC18 using T4 DNA Ligase (Thermo Scientific). |
</li> | </li> | ||
</ul> | </ul> | ||
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<b>Changing the <i>Xba</i>I site in the <i>tar</i> sequence via QuikChange</b><br> | <b>Changing the <i>Xba</i>I site in the <i>tar</i> sequence via QuikChange</b><br> | ||
<ul> | <ul> | ||
- | <li>Experiment: <br>DNA was prepped using peqGOLD | + | <li>Experiment: <br>DNA was prepped using peqGOLD Miniprep Kit (Peqlab). A DNA test digest with <i>Eco</i>RI and <i>Xba</i>I was performed according to <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a> and analyzed on a 1% agarose gel. Test digest showed 8 out of 10 clones contained the construct pUC18_TAR. QuikChange PCR was performed using designed primers and <i>Pfu</i> Turbo polymerase (see QuikChange protocol <a href="https://2012.igem.org/Team:Goettingen/Project/Methods#QuikChange_Protocol">here</a>). Resulting construct was labeled pUC18_TAR_QC. |
</li> | </li> | ||
</ul> | </ul> |
Latest revision as of 11:24, 22 September 2012
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#3 Chemoreceptor Library - 5th WeekBack to overview
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