Team:Bielefeld-Germany/Labjournal/week16

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==Week 16 (08/13 - 08/19/12)==
 
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===Monday August 13th===
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* '''Team Cultivation & Purification:'''
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** Today we discussed that we may get activity if we start to purify our laccases. So from now we will purify our products before measuring them. We hope this will bring promising results.
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===Tuesday August 14th===
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* '''Team Arabidopsis Laccase''': Today our cDNA took a bath in ethanol and got cleaned. We are pretty sure our next PCR will be a lot more successful. Check protocols for further information about the cDNA washing via [https://2012.igem.org/Team:Bielefeld-Germany/Protocols/Genetics#Ethanol_precipitation_to_clean_DNA ethanol precipitation].
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* '''Team Cultivation & Purification:'''
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** Made preculture of ''E.coli'' KRX without plasmid and with plasmids containing laccases from ''B.halodurans'', ''E.coli'', ''B.pumilus'', ''T.thermophilus'' and ''X.campestris'' as well as [http://partsregistry.org/Part:BBa_K525710 BBa_K525710 (Ligase A)].
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===Wednesday August 15th===
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<!-- navigator -->
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*'''Team Wiki''': We set up some more wiki rules. Today´s rules were about citations. We agreed von some standards that will for sure help make the wiki a little prettier. Have a look:
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** <span style="text-decoration: underline;">Paper</span>: Exampleman M ''et al''. (2002). The example paper. The example journal (Volume): pages.
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** <span style="text-decoration: underline;">Website</span>: Name of website, URL, Index, date site visited
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<ul style="list-style-type:none">
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** <span style="text-decoration: underline;">Book</span>: Examplewomen M ''et al''. (1999). The example book. The example publisher (edition).
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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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Next will be a standard for charts.
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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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* '''Team Cultivation & Purification:'''
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</ul>
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** flask cutivation of ''E.coli'' KRX with laccases from ''B.halodurans'', ''E.coli'', ''B.pumilus'', ''T.thermophilus'' and ''X.campestris''. As positive control we used ''E.coli'' KRX with [http://partsregistry.org/Part:BBa_K525710 BBa_K525710 (Ligase A)].
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::--> Settings: 1L flasks without baffles, final volume 250mL, autoinduction medium supplemented with 60µg/mL chloramphenicol, 37°C, 120rpm, single determination
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===Thursday August 16th===
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===Friday August 17th===
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===Saturday August 18th===
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* '''Team bacterial laccases''': Today we performed a ligation from the digestions we have done before. The ligation included of course the T7 promotor, the backbone pSB1C3 and the individual laccase genes. Ligation was performed with T4 ligase for 20 minutes at room temperature and stopped through a five minute incubation at 70°C.
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===Sunday August 19th===
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==Week 16 (08/13 - 08/19/12)==
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* '''Team Site Directed Mutagenesis:'''
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Plasmidisolation of X.camp_2247- 1-4 and _3633 1-4
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Test-restriktion of  X.camp_2247 1-4 and _3633 1-4 with PstI for 3 hours
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2247:1,2+4 no Mutation
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3633:2+3 no Mutation.
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2247-3 and 3633-1 only cut once (should be twice)
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Picked 4 new colonys per site and plated the on CM-agar
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Colony-PCR of 10 Colonies of T10
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1,2,3,4 bands at 1100 - should be RFP
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9 + 10 bands could be higher but not 1600 as needed… plated anyway for Test-restriktion.
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{{Team:Bielefeld/Sponsoren}}
{{Team:Bielefeld/Sponsoren}}

Latest revision as of 21:26, 25 September 2012


Week 16 (08/13 - 08/19/12)

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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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