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Team:UPO-Sevilla/Notebook/07 15 - Revision history
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Lepavgom at 11:56, 24 October 2010
2010-10-24T11:56:50Z
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Lepavgom
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Lepavgom at 18:13, 17 October 2010
2010-10-17T18:13:54Z
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Lepavgom
http://2010.igem.org/wiki/index.php?title=Team:UPO-Sevilla/Notebook/07_15&diff=99481&oldid=prev
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2010-10-13T16:13:09Z
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<h1>July, 15th</h1><br />
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<h2>Production Team</h2><br />
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<p><strong>Paola Gallardo.</strong> Using the ligation products we got the last day, we transformed competent bacteria and spread in LB+Cm plates once again. We hoped to get better results this time. New inocula of UPO16 (aspartate ammonia lyase) and UPO3 (rrnBT1-T7TE) were set up.</p><br />
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<p><strong>David Caballero.</strong> We continued without getting all parts, so we performed other PCR reactions for parts still not amplified. PCR reactions products were analyzed by 0.8% agarose gel electrophoresis and we realized presence of parts <i>fecR*, </i>P<i>fecA, gltD**</i> and <i>fecI-fecR*</i>. How it showed, the biggest parts, <i>fecA*</i> (2,3 kbp) and <i>gltB</i> (4,5 kbp), could not be obtained by PCR cycle we used.</p><br />
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<h2>Assembly Team</h2><br />
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<p>Today we’ve started with the UPO16+3 assemblies again.</p><br />
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<p>After yesterday’s disappointment, we started over again digesting and ligating UPO 16+UPO 3. The GINGO kit arrived this morning, that’s why we hope to make ligation faster.</p><br />
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<li>Digestion and ligation of UPO3 and UPO16 using the GINGO kit. Later, transformation in DH5α</li><br />
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<li><i>E.Coli</i> transformation with UPO2+13 and UPO1+19.</li><br />
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<li>Colony PCR of bacteria which have been growing all night</li><br />
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Lepavgom