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Team:UPO-Sevilla/Notebook/07 26
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<h2>Production Team</h2> | <h2>Production Team</h2> | ||
| - | <p><strong>Paola Gallardo.</strong> Products of the first PCR reaction were analyzed by electrophoresis (0'8 %). We got four bands clearly-defined with the correct lengths ( | + | <p><strong>Paola Gallardo.</strong> Products of the first PCR reaction were analyzed by electrophoresis (0'8 %). We got four bands clearly-defined with the correct lengths (635bp and 732bp for <i>gltD</i>, 1206bp and 312bp for <i>fecI-fecR</i>). Purification from gel.</p> |
<p><strong>David Caballero.</strong> Plate analysis: colonies were presented in all plates except for <i>fecA*</i> and control plates. We verified the presence of <i>fecI</i> and P<i>fecA</i> parts in colonies by colony PCR reactions and 1.5% agarose gel electrophoresis analysis. In other way, to obtain <i>fecA*</i> part we digested by restriction enzymes the amplified by PCR part again, ligated it and transformed competent bacteria which remained overnight at 37ºC in LB+Km plate.</p> | <p><strong>David Caballero.</strong> Plate analysis: colonies were presented in all plates except for <i>fecA*</i> and control plates. We verified the presence of <i>fecI</i> and P<i>fecA</i> parts in colonies by colony PCR reactions and 1.5% agarose gel electrophoresis analysis. In other way, to obtain <i>fecA*</i> part we digested by restriction enzymes the amplified by PCR part again, ligated it and transformed competent bacteria which remained overnight at 37ºC in LB+Km plate.</p> | ||
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<p>And again, digestion and ligation of UPO1 and UPO2 using the GINGO kit. Later, transformation in DH5α</p> | <p>And again, digestion and ligation of UPO1 and UPO2 using the GINGO kit. Later, transformation in DH5α</p> | ||
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| + | <a class="return_button" href="/Team:UPO-Sevilla/Notebook/07_23" title="Go to 23rd of July"><span>Go to 23rd of July</span></a> | ||
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| + | <a class="next_button" href="/Team:UPO-Sevilla/Notebook/07_27" title="Go to 27th of July"><span>Go to 27th of July</span></a> | ||
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<a class="return_button" href="/Team:UPO-Sevilla/Notebook" title="Notebook"><span>Return to Notebook</span></a> | <a class="return_button" href="/Team:UPO-Sevilla/Notebook" title="Notebook"><span>Return to Notebook</span></a> | ||
Latest revision as of 21:51, 26 October 2010
July, 26th
Production Team
Paola Gallardo. Products of the first PCR reaction were analyzed by electrophoresis (0'8 %). We got four bands clearly-defined with the correct lengths (635bp and 732bp for gltD, 1206bp and 312bp for fecI-fecR). Purification from gel.
David Caballero. Plate analysis: colonies were presented in all plates except for fecA* and control plates. We verified the presence of fecI and PfecA parts in colonies by colony PCR reactions and 1.5% agarose gel electrophoresis analysis. In other way, to obtain fecA* part we digested by restriction enzymes the amplified by PCR part again, ligated it and transformed competent bacteria which remained overnight at 37ºC in LB+Km plate.
Assembly Team
We repeated PCR colony reaction because we hadn’t anything confirmed yet. We analyzed UPO1+UPO2 with Nhe1 but results were negative. Moreover, we did PCR colony reaction for the new UPO1+2 part. We are moving forward too slow. In addition we are waiting for the synthesized DNA... We hope it comes soon!
And again, digestion and ligation of UPO1 and UPO2 using the GINGO kit. Later, transformation in DH5α
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