Team:Bielefeld-Germany/Test

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{{Team:Bielefeld/Head}}
{{Team:Bielefeld/Head}}
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==Week 24 (10/08 - 10/14/12)==
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<a href="http://2012.igem.org/Team:Bielefeld-Germany/Public_relations_Overview#8"><img src="http://2012.igem-bielefeld.de/includes/wiki/images/Pfeil_links2.png"></a>
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        <div id=page-title>
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            <span id=page-title-text>
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                How our project reaches the world
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            </span>
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        </div>
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  <div id="grey_bg">
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<script src="http://2012.igem-bielefeld.de/includes/wiki/flex/js/jquery.flexslider.js"></script>
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===Monday October 08th===
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All:
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<script type="text/javascript" charset="utf-8">
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Day off in Amsterdam!!!
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===Tuesday October 09th===
 
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===Wednesday October 10th===
 
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* '''Team Shuttle Vector:'''
 
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** Ligation of pSB1C3::BBa_K863202 construct in KRX cells was done.
 
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* '''Team Cellulose Binding Domain:'''
 
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** Gradient-PCRs of CBDcex_Freiburg; CBDclos_Freiburg; GFP_Freiburg and ecol_Freiburg (47 °C to 67°C)
 
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*** Product:
 
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**** CBDcex_Freiburg at all temperatures
 
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**** CBDclos_Freiburg at all temperatures
 
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**** GFP_Freiburg at all temperatures
 
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**** ecol_Freiburg at 53 °C to 61°C (best at 57 to 59 °C)
 
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*** pooled fractions for gel clean up
 
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* '''Team Site Directed Mutagenesis:'''
 
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** Ordered Primers for the SDM of the illegal XbaI restriction site in the shuttle vector
 
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* '''Team Cultivation and Purification:'''
 
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**Made precultures of ''E.&nbsp;coli'' KRX with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863000 BBa_K863000] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863005 BBa_K863005]for 19L fermentation (500mL preculture)
 
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===Thursday October 11th===
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<!-- Slider Container and Images -->
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* '''Team Cellulose Binding Domain:'''
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<div style="padding:5px; z-index:1;">
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** Clean-Up of the Gel-pieces (from PCR-products)
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        <div class="flexslider" style="margin-top:10px;">
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** Restriction of PCR-products (3h):
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            <ul class="slides">
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*** GFP_Freiburg:
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                <li>
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**** XbaI+PstI Tango-Buffer for Ligation
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                    <div class="frontpage-slide">
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**** XbaI+AgeI Tango 4xAgeI for Assembly
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                        <img src=""></a>
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*** CBDclos/cex:
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                        <div class="slide-right-col">
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**** NgoMIV+PstI P.4+BSA 2xPstI for Assembly
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** Assembly and Ligation (1h with T4):
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*** GFP_Freiburg + CBDcex_Freiburg + J61101
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*** GFP_Freiburg + CBDclos_Freiburg + J61101
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*** GFP_Freiburg + J61101
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** Transformation in ''E. coli'' KRX (3 µL, chemical):
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*** GFP_Freiburg + CBDcex_Freiburg + J61101
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*** GFP_Freiburg + CBDclos_Freiburg + J61101
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*** GFP_Freiburg + J61101
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** All Transformations were plated on AMP-select-agar
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*'''Team Cultivation and Purification'''
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** 12 L Cultivation of ''E.&nbsp;coli'' KRX with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863000 BBa_K863000] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863005 BBa_K863005]
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***Settings:
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Bioengineering NFL 19L fermenter, autoinduction HSG medium, 60 µg/mL chloramphenicol, 19 L, 37 °C, stirrer on cascade to hold a pO2 of 50 %, airflow: 12 NL/m, 16 hours. HSG medium was choosen to get a high biomass concentration with hope for a higher amount of laccases.
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**Made precultures of ''E.&nbsp;coli'' Rosetta Gami 2 containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863012 BBa_K863012](09/09) and <partinfo>BBa_K863022</partinfo> (09/09) behind a constitutive promoter. (500mL preculture)
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                        </div>
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                        <div class=clear></div>
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                    </div>
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                </li>
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                <li>
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                    <div class="frontpage-slide">
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                        <img src=""></a>
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                        <div class="slide-right-col">
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===Friday October 12th===
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                        </div>
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* '''Team Cellulose Binding Domain:'''
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                        <div class=clear></div>
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** A lot of colonies on all three dishes (J61101+GFP_Freibug;J61101+GFP_Freibug+CBDcex;J61101+GFP_Freibug+CBDclos)
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                    </div>
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*** no obvious green fluorescence, not even at J61101+GFP
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                </li>
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*** Maybe the combination of J23100 and J61101 isn't strong enough to express a visible amount of GFP
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                <li>
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** Colony-PCR of 16 colonies of every dish
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                    <div class="frontpage-slide">
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*** J61101 + GFP_Freiburg:
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                        <img src=""></a>
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**** 16/16 positive
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                        <div class="slide-right-col">
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**** plated two on select-agar
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*** J61101 + GFP_Freiburg + CBDcex:
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**** 8/16 positve
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**** plated three on select-agar
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*** J61101 + GFP_Freiburg + CBDclos:
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**** 7/16 positive
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**** plated three on select-agar
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* '''Team Shuttle Vector:'''
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** The induction with 0.5% (v/v) methanol of P. pastoris GS115 cells with integrated BBa_K863204::GFP was started.
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* '''Team Substrate Analysis''':
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** Since we have seen some new peaks after Estradiol treated with Laccases in the LC-MS, we wanted to have more degradation products to do an MS so we used a higher concentradion of Estradiol and Laccase and let them incubate for 66 hours.
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* '''Team Cultivation and Purification'''
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                        </div>
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** 12L Cutlivations of ''E.&nbsp;coli'' KRX with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863000 BBa_K863000] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863005 BBa_K863005] were harvested and stored at 4°C until purification.
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                        <div class=clear></div>
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** 12 L Cultivation of ''E.&nbsp;coli'' Rosetta Gami 2 containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863012 BBa_K863012](09/09) and <partinfo>BBa_K863022</partinfo> (09/09)
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                    </div>
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*** Settings:
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                </li>
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Bioengineering NFL 19L fermenter, HSG medium, 60 µg/mL chloramphenicol, 300 µg/mL ampicillin, 12 L, 37 °C, stirrer on cascade to hold a pO2 of 50 %, airflow: 12 NL/m, 22-24 hours. HSG medium was chosen to get a high biomass concentration with hope for a higher amount of laccases.
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                <li>
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                    <div class="frontpage-slide">
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                        <img src=""></a>
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                        <div class="slide-right-col">
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===Saturday October 13th===
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                        </div>
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* '''Team Shuttle Vector:'''
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                        <div class=clear></div>
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** The P. pastoris GS115 cells integrated with BBa_K863204::GFP were also inducted with 0.5% (v/v) methanol (100%).
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                    </div>
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                </li>
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                <li>
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                    <div class="frontpage-slide">
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                        <img src=""></a>
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                        <div class="slide-right-col">
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* '''Team Cultivation and Purification'''
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** 12L Cutlivations of ''E.&nbsp;coli'' Rosetta Gami 2 containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863012 BBa_K863012](09/09) and <partinfo>BBa_K863022</partinfo> (09/09) were harvested and stored at 4°C until purification.
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</div>
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                        <div class=clear></div>
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</div>
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                </li>
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            </ul>
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        </div>
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===Sunday October 14th===
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    </div>
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* '''Team Shuttle Vector''':
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** The P. pastoris GS115 cells integrated with BBa_K863204::GFP were also inducted with 0.5% (v/v) methanol (100%).
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* '''Team Fungal and Plant Laccases:'''
 
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** Plasmid isolation of pSB1C3::BBa_K863204::TV5 for transformation into P. pastoris GS115 was done. After isolation the plasmid was digested with [[Team:Bielefeld-Germany/Protocols/Materials#Used_enzymes |NotI]], purified with the PCR clean up kit and transformed via [[Team:Bielefeld-Germany/Protocols/molecular_genetics#Transformation_of_yeast_cells |protocol]] in P. pastoris GS115.
 
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* '''Team Cellulose Binding Domain:'''
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<ul>
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** Tried to isolate all plates, but only got low concentrations (5 ng/µL - 20 ng/µL)
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** plated eight different colonies for isolation
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* '''Team Substrate Analysis''':
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<li><b>We did it! About our qualification for the World Championship in Boston</b><br><br>
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** We took sample from the reaction of the 12th October.
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&#10158; <a href="http://www.laborwelt.de/aktuelles/nachrichten/2012-q4/igem-fuenf-teams-nach-boston.html">iGEM: Fünf Teams nach Boston, Laborwelt - 10/09/12</a><br>
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&#10158; <a href="http://www.biotechnologie.de/BIO/Navigation/DE/root,did=154774.html?listBlId=74462&">iGEM: Fünf Teams lösen Ticket für Boston, biotechnologie.de - 10/10/12</a></li><br>
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<li><b>Neue Züricher Zeitung</b><br><br>
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&#10158; <a href="http://www.nzz.ch/wissen/wissenschaft/arzneimittel-in-gewaessern-minimieren-1.17368069">Arzneimittel in Gewässern minimieren - 07/18/12</a></li>
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<br>
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<li><b>Neue Westfälische</b><br><br>
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&#10158; <a href="http://2012.igem.org/wiki/images/7/7e/Article_NW.png">Das Wasser wird weiblich - Article in our local newspaper Neue Westfälische, Bielefelder Tageblatt, No. 160, Author: Uwe Pollmeier - 07/12/12</a></li>
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<br>
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<li><b>LIVE radio interview at Radio Hertz</b><br><br>
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&#10158; <a href="http://www.radiohertz.de/beta-site/livestream/">07/04/2012 at 6pm</a></li>
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<br>
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<li><b>WDR5 Leonardo</b><br><br>
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&#10158; <a href="http://www.wdr5.de/sendungen/leonardo/s/d/06.09.2012-16.05/b/hormone-und-betablocker-im-gewaesser.html">09/06/2012 at 4pm</a></li>
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<br>
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<li><b>Our project reaches the world </b><br><br>
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&#10158; <a href="http://www.wired.co.uk/news/archive/2012-06/25/tree-fungus-oestrogen-filter">Great Britain - Wired.co.uk - 06/25/2012</a><br>
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&#10158; <a href="http://scienceblog.com/55186/filtering-estrogen-from-drinking-water/?utm_source=feedburner&utm_medium=feed&utm_campaign=Feed%3A+scienceblogrssfeed+%28ScienceBlog.com%29">Scienceblog - 06/25/2012</a><br>
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&#10158; <a href="http://www.news-medical.net/news/20120626/Biological-filter-to-remove-estrogens-from-waste-water-and-drinking-water.aspx">Australia - News Medical - 06/26/2012</a><br>
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&#10158; <a href="http://savethewater.org/tag/bielefeld/">Los Angeles, USA - Save the Water<sup>TM</sup> - 06/26/2012</a><br>
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&#10158; <a href="http://www.eurekalert.org/pub_releases/2012-06/uob-ref062512.php">Washington, DC - EurekAlert! - 06/25/2012</a><br>
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&#10158; <a href="http://savethewater.org/tag/bielefeld/">Hollywood, USA - Save the water - 06/26/2012</a></li><br>
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&#10158; <a href="http://www.biofuelsdigest.com/biobased/2012/06/27/bielefeld-university-students-participating-in-igem-competition/">Miami, USA - BioBasedDigest - 06/27/2012</a></li><br>
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&#10158; <a href="http://phys.org/news/2012-06-estrogen.html"> Phys.org - 06/25/2012</a><br>
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&#10158; <a href="http://www.redorbit.com/news/science/1112645127/drinking-water-minus-the-estrogen/">Texas - RedOrbit - 06/25/2012</a><br>
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&#10158; <a href="http://www.provital-blog.com/en/nutrition/drinks/biological-filter-to-remove-estrogens-from-waste-water-and-drinking-water/">Provital-blog.com - 06/30/2012</a><br>
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&#10158; <a href="http://www.innovations-report.com/html/reports/awards_funding/removing_estrogen_drinking_water_197866.html"> Schmitten, Germany - innovations report - 06/25/2012</a><br>
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&#10158; <a href="http://www.pressrelations.de/new%5C/standard/result_main.cfm?r=499469&sid=&aktion=jour_pm&quelle=0&n_firmanr_=102898&pfach=1&detail=1&sektor=pm&popup_vorschau=0"> Düsseldorf, Germany - pressrelations - 06/25/2012</a><br>
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&#10158; <a href="http://www.sciencecodex.com/removing_estrogen_from_drinking_water-93912">Science Codex - 06/25/2012</a><br>
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&#10158; <a href="http://healthresearchreport.me/2012/07/01/132nd-issue-health-technology-research-synopsis-29-jun-2012/">Health Research Report - 06/29/2012</a><br>
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&#10158; <a href="http://www.scoop.it/t/syntheticbio/p/2037868685/removing-estrogen-from-drinking-water">Scoop.it! - 06/26/2012</a><br>
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&#10158; <a href="http://www.medindia.net/news/estrogen-removal-from-drinking-water-using-biological-filters-103306-1.htm?utm_source=feedburner&utm_medium=feed&utm_campaign=Feed%3A+allhealthnews+%28Medindia+Health+News%29">Chennai, India - MedINDIA Network for Health - 06/27/2012</a><br>
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&#10158; <a href="http://www.analytic-news.com/press/2012/69.html">Ober-Ramstadt, Germany - News Analytic - 06/26/2012</a><br>
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<li><b>Removing estrogen from drinking water</b><br><br>
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&#10158; <a href="http://idw-online.de/pages/de/news484982">Informationsdienst Wissenschaft - 06/25/2012</a><br>
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&#10158; <a href="http://www.bio.nrw.de/nachrichten">BIO.NRW - 06/25/2012</a><br>
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&#10158; <a href="http://www.laborpraxis.vogel.de/forschung-und-entwicklung/analytik/articles/369125/">Laborpraxis.de - 06/25/2012</a>
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<br>
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&#10158; <a href="http://www.uni-protokolle.de/nachrichten/id/240229/">uni-protokolle.de - 06/25/12</a><br>
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&#10158; <a href="http://mag3.i-magazine.de/magPage/gwf_Wasser_Abwasser_Anlagen_und_Verfahren_zur_Wasseraufbereitung/Seite_28/#/28"> gwf Wasser Abwasser - 7-8/2012 p. 766-767</a></li><br>
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&#10158; <a href="http://www.bio.nrw.de/en/latest_news"> BioNRW </a></li><br>
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<li><b>Students from Bielefeld University are participating in the international iGEM competition</b><br><br>
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&#10158; <a href="http://ekvv.uni-bielefeld.de/blog/pressemitteilungen/entry/%C3%B6strogen_aus_trinkwasser_entfernen_nr">Bielefeld University - 06/25/2012 - German</a><br>
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&#10158; <a href="http://ekvv.uni-bielefeld.de/blog/uninews/entry/removing_estrogen_from_drinking_water">Bielefeld University - 06/25/2012 - English</a><br>
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&#10158; <a href="http://www.laborjournal.de/rubric/aktuell/index.lasso">Synthetische Biologie: Das Bielefelder iGEM-Team, Laborjournal - 10/2012</a></li><br>
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<li><b>Street Science</b><br><br>
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&#10158; <a href="http://www.bielefeld.de/de/pressedienst/detail.html?id=2012-08-21-12.20.37.146281"> Press release for our Street Science Day - 06/25/2012</a><br>
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&#10158; <a href="http://2012.igem.org/wiki/images/b/bf/Street_Science_NW_2708.jpg">DNA isolieren? Das kann doch jeder! (Street Science) - Article in our local newspaper Westfalen-Blatt - 08/27/12</a><br>
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&#10158; <a href="http://2012.igem.org/wiki/images/f/fe/Street_Science_NW_2708_2.jpg">Spielerische Wissenschaft (Street Science) - Article in our local newspaper Neue Westfälische, Bielefelder Tageblatt, No. 199, Author: Jonny Dähne - 08/27/12</a></li><br>
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</ul>
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{{Team:Bielefeld/Sponsoren}}
{{Team:Bielefeld/Sponsoren}}

Latest revision as of 20:20, 26 October 2012

How our project reaches the world


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