Team:Bielefeld-Germany/Labjournal/week2

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=Labjournal=
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week1 Week1]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week2 Week2]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week3 Week3]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week4 Week4]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week5 Week5]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week6 Week6]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week7 Week7]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week8 Week8]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week9 Week9]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week10 Week10]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week11 Week11]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week12 Week12]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week13 Week13]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week14 Week14]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week15 Week15]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week16 Week16]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week17 Week17]
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[https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week18 Week18]
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==Week 2 (05/07 - 05/13/12)==
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<ul style="list-style-type:none">
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/Starting"><strong>Prologue</strong></a></li>
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                <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week1"><strong>Week 1</strong></a></li>
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                <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week2"><strong>Week 2</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week3"><strong>Week 3</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week4"><strong>Week 4</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week5"><strong>Week 5</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week6"><strong>Week 6</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week7"><strong>Week 7</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week8"><strong>Week 8</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week9"><strong>Week 9</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week10"><strong>Week 10</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week11"><strong>Week 11</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week12"><strong>Week 12</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week13"><strong>Week 13</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week14"><strong>Week 14</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week15"><strong>Week 15</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week16"><strong>Week 16</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week17"><strong>Week 17</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week18"><strong>Week 18</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week19"><strong>Week 19</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week20"><strong>Week 20</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week21"><strong>Week 21</strong></a></li>
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<li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week22"><strong>Week 22</strong></a></li>
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* Primerdesign for isolation of laccases from genomic DNA of ''Xanthomonas campestris B100'', ''E. coli BL21(DE3)'' and ''Bacillus pumilus ATCC7061''. The forward primers were designed with T7 promotor-overhanging ends after prefix and the first 20 bases of the wanted gene. The reverse primers were designed with a HIS-Tag and tweo stop codons before suffix and the last 20 bases of the wanted gene without the stop codon.
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* Successful PCRs of laccase genes CopA from ''Xanthomonas campestris B100'', CueO from ''E. coli BL21(DE3)'' with the isolated genomic DNA as template.
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* Because we want to characterise laccases from different bacteria we had to order the bacterial strains, which weren't available at the University Bielefeld, from [http://www.dsmz.de/|''DSMZ''].
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==Week 2 (05/07 - 05/13/12)==
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* [http://www.dsmz.de/catalogues/details/culture/DSM-7039.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|''Thermus thermophilus HB27'']
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* [http://www.dsmz.de/catalogues/details/culture/DSM-18197.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|''Bacillus halodurans C-125'']
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* [http://www.dsmz.de/catalogues/details/culture/DSM-40069.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|''Streptomyces lavendulae REN-7'']
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* [http://www.dsmz.de/catalogues/details/culture/DSM-40236.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|''Streptomyces griseus IFO 13350'']
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* After some empty agarose gels we finally isolated  laccase gene CotA from ''Bacillus pumilus ATCC7061'' as a PCR product. As template we used the plasmid we got from the Swiss working group.
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* '''Team Modeling''': Looking for suitable Software and enzymkinetics to model the degradation of our substartes with the different laccases. Finding the Michaelis-Menten kinetics and matlab.
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=== Monday May 7th ===
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* '''Team Student Academy:''' First transformation of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 BBa_I13522] and plating on selective agar
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** Electroporation setup: U= 2,5 kV C= 25 µF and R= 400 &Omega;
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{| class="wikitable"
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! Material !! Volume
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| ''E. coli'' KRX competent cells || 50 µL
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| glycerol (10 %) || 50 µL
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| plasmid || 1 µL
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** Result: We got little colonies. There weren’t any green colonies and only very pale red fluorescent colonies.
 
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=== Tuesday May 8th ===
 
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* '''Team Student Academy:''' Repetition of the transformation didn’t change the result. We made a liquid culture of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450], but it did not fluoresce. Searching for mistakes and alternatives. Maybe competent cells are not that good and in case of RFP the lacI sensitivity could be the problem.
 
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=== Wednesday May 9th ===
 
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=== Thursday May 10th ===
 
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* '''Team Student Academy''' Testing the competent cells by transformation of pUC19. The transformation did not work that good, so that we produced new ones.
 
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=== Friday May 11th ===
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{{Team:Bielefeld/Sponsoren}}

Latest revision as of 21:10, 25 September 2012

Week 2 (05/07 - 05/13/12)

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About our Wiki

Fifteen students from different fields of study from the Bielefeld University aspired to complete a project called TOXIC COMPOUNDS IN NATURAL WATER - A CASE FOR LACCASE. In his Wiki we document our achievements and our approach.

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