Team:Bielefeld-Germany/Labjournal/week13

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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 13 (07/23 - 07/29/12)==
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===Monday July 23rd===
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* Some of our team members were at CAS conference in Munich from 07/23 - 07/25/12.
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* Successful PCRs of laccase gene lac5 from ''Trametes versicolor'' with plasmid DNA from Uni Greifswald as template.
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* '''Team Site Directed Mutagenesis:''' Transfered 4 colonys per petri dish to an new one for plasmidisolation
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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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===Tuesday July 24th===
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* '''Team Activity Tests:''' Today was the moment of truth. We received the first recombinant laccases from the cultivation team. We started with testing the supernatant from the cultivated cells to check whether they secrete laccase. We added 0,1 mM ABTS in each well that was filled with supernatant from ''Escherichia coli'', ''Bacillus pumilus'', ''Bacillus halodurans C-125'', ''Xanthomonas campestris pv. campestris B100'' and ''E. coli KRX'' (negative control - cells without plasmid) and measured as usual. There was no change in the OD so that we assume that no secretion takes place. Afterwards we tested potential laccase activity by filling each well with laccase and buffer (check protocols to see which buffer we used) and added 0,1 mM ABTS. Unfortunately no activity was seen. We will try again soon.
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* '''Team Site Directed Mutagenesis:''' Plasmidisolation of B.pumilus 2883(I-IV); X.campestris 2247(I-IV); T.thermophilus 2796(I-IV) and E.coli 2307 (III).
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==Week 13 (07/23 - 07/29/12)==
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===Wednesday July 25th===
 
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* '''Team Site Directed Mutagenesis:''' Testrestriktion of the Plasmids revealed that ''B.pumilus'' III;IV; ''E.coli'' III;''T.thermophilus''.I;II seem to be mutated corrcetly but ''X.campestris'' is untouched (no Band at 3636).
 
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**SDM-PCR with: ''B.pumilus'' III(III)+IV(IV) with the SDM-primermix to mutate bp 2317
 
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started a SDM-PCR with ''X.campestris'': with 2247-primers again and another one with the primermix to mutate 3633
 
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===Thursday July 26th===
 
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* '''Team Modeling''' and '''Team Sponsoring''': meeting Mr. ??? from the clarification plant of Schloß Holte and finding a new cooperation partner. He want to give us the information we need to equate our model and design our cleaner. On top he wants to ask if the clarification plant can sponsor our project.
 
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===Friday July 27th===
 
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===Saturday July 28th===
 
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* '''Team Activity Tests:''' See, we are back already! Today we received different samples from ''Escherichia coli'', ''Bacillus pumilus'', ''Bacillus halodurans C-125'', ''Thermus thermophilus'', ''Xanthomonas campestris pv. campestris B100'' and ''E. coli KRX'' (negative control - cells without plasmid). The cells were all cultivated at 30°C and supplied with copper. We tested each one of them but none of them showed laccase activity.
 
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===Sunday July 29th===
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{{Team:Bielefeld/Sponsoren}}

Latest revision as of 21:23, 25 September 2012


Week 13 (07/23 - 07/29/12)

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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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