Team:Bielefeld-Germany/Labjournal/week1

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{{Team:Bielefeld/Head}}
{{Team:Bielefeld/Head}}
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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 1 (04/30 - 05/06/12)==
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* Start of our WET LAB time.
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* Generating new competent ''E.coli KRX cells''.
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<div id="nav" class="tabs">
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* Cultiviation of ''Xanthomonas campestris B100'' and ''E. coli BL21(DE3)''. The bacterial strains we got from a working group at our University. After cultivation we isolated the genomic DNA. The DNA was needed as template for PCRs to purify the wanted laccase ORFs.  
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* Sending requests to working groups for different plasmids , which have already worked with laccases and described them in their papers. Unfortunately just one answer came back (thanks a lot to them).So we got a vector with the laccase-ORF CotA from ''Bacillus pumilus ATCC7061'' from the Swiss Federal Laboratories for Materials Science and Technology, Laboratory for Biomaterials in Switzerland.
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<ul style="list-style-type:none">
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=== Thursday May 3th ===
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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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* '''Team Bacterial Laccase''': After the vector has arrived, we transformed it into the competent ''E.coli KRX'' which we have already generated. The protocol we used was as followed:  
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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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** The electroporation setup: U= 2,5kV C= 25 µF and R= 400 <math>\omega</math>
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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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** Since we did not know the efficient of our competent KRX we used two different ''E.coli'' volumes for the transformation, 50µL and 100µL. We gave 50µL 10% Gylcerol to the reaction tubes with 1µL of the vector DNA (Bacillus pumilus). After the transformation we plated them into ampicillin plates.
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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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* '''Team Bacterial Laccase''': PCR with the ''Xanthomonas campestris B100'' and ''E. coli BL21(DE3)'' genomic DNA.  
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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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'''PCR table'''
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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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{| class="wikitable"
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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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|-
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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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! Material !! Volume
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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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|-
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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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| Buffer (10x Phusion) || 10µL
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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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|-
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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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| Phusion Polymerase || 0,5µL
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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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|-
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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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| dNTPs || 1µL
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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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|-
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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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| Primer Mix || 1µL
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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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|-
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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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| Template DNA || 1µL
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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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|-
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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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| DMSO || 1,5µL
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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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|-
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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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| Water || 35µL
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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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|}
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''' PCR program'''
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{| class="wikitable"
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! Temperature !! Time
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| 1) 98°C || 30 sec
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| 2) 98°C || 15 sec
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| 3) 62°C || 45 sec
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| 4) 72°C || 1 min
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| 5) 72°C || 3 min
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| 6) 12°C ||
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|}
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Cycle between step 2 and 4 35 times.
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weekly seminar:
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</div>
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* Do we want to order strains of ''Trametes versicolor'' and ''Trametes villosa''?
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</div>
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* Gathering information about signal sequences in yeast
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</html>
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* Decision to create a database, so that we can easily number and inscribe our lab results
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* Decision to arrange a summer school for pupils in their last year before the final exams
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<div style="text-align:justify;">
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* Discussion about how to meet a member of the german ''Bundestag'' (the german parliament)
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{{Team:Bielefeld/Sponsoren}}

Latest revision as of 21:08, 25 September 2012



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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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