Team:HKU HongKong/Data/Weekly Notebook.html

From 2012.igem.org

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   </span><span style="font-size: 10pt; font-family: Tahoma">2)</span><font color="#232323" size="2" face="Tahoma"><b>Test digestion of amplified pvdQ with EcoRI only, EcoRI & SpeI, PstI, and EcoRI & XbaI. </b></font></h2>
   </span><span style="font-size: 10pt; font-family: Tahoma">2)</span><font color="#232323" size="2" face="Tahoma"><b>Test digestion of amplified pvdQ with EcoRI only, EcoRI & SpeI, PstI, and EcoRI & XbaI. </b></font></h2>
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  <li><em>Result: Two bands present in the EcoRI only digestion instead of the  expected single, linear fragment. Indicates presence of a skeptical EcoRI site  – a sequence that is not identical to the conventional EcoRI palindromic  sequence but similar such that EcoRI restriction enzyme non-specifically cuts  it). </em></li>
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</span><span style="font-size: 10pt; font-family: Tahoma">3)</span><font color="#232323" size="2" face="Tahoma"><b> Repeat the digestion of pvdQ with EcoRI and SpeI multiple times to obtain the desirable conditions of the digestion to prevent Star Activity. </b></font></h2>
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  <font size="2" face="Tahoma" color="#232323">&nbsp;</font><font color="#232323"><span style="font-size: 10pt; font-family: Tahoma"><br>
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</span><b><font size="2" face="Tahoma">3)
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Repeat the digestion of pvdQ with EcoRI and SpeI multiple times to obtain the desirable conditions of the digestion to prevent Star Activity.</font></b></font></p>
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<font size="2" face="Tahoma" color="#232323">4)</font> Mass single digestion of the amplified pvdQ with EcoRI.  Desirable fragment size should be about 5,600bp. The fragments that result due to Star activity are 3,800 and 1,800bp. </b></p>
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<font size="2" face="Tahoma" color="#232323">4)</font>
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5). Gel purification and Nano-drop concentration check of digested pvdQ.
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Revision as of 01:42, 27 September 2012

Team:HKU Hong Kong - 2012

Team:HKU HK

From 2011.igem.org