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| | {{Team:Bielefeld/Head}} | | {{Team:Bielefeld/Head}} |
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| - | <div style="text-align:justify;">
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| - | =Labjournal=
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| - | [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week1 Week1]
| + | <html> |
| - | [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week2 Week2]
| + | <style type="text/css"> |
| - | [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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| - | [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week19 Week19]
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| - | [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week20 Week20]
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| - | [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week21 Week21]
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| | + | ul {list-style-image:none;} |
| | + | #bodyContent{ |
| | + | background-color: white; |
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| - | ==Week 5 (05/28 - 06/03/12)==
| + | </style> |
| | | | |
| - | ===Monday May 28th=== | + | <!-- navigator --> |
| | + | <div id="nav" class="tabs"> |
| | + | <div class="scroller"> |
| | + | <ul style="list-style-type:none"> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/Starting"><strong>Prologue</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week1"><strong>Week 1</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week2"><strong>Week 2</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week3"><strong>Week 3</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week4"><strong>Week 4</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week5"><strong>Week 5</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week6"><strong>Week 6</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week7"><strong>Week 7</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week8"><strong>Week 8</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week9"><strong>Week 9</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week10"><strong>Week 10</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week11"><strong>Week 11</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week12"><strong>Week 12</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week13"><strong>Week 13</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week14"><strong>Week 14</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week15"><strong>Week 15</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week16"><strong>Week 16</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week17"><strong>Week 17</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week18"><strong>Week 18</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week19"><strong>Week 19</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week20"><strong>Week 20</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week21"><strong>Week 21</strong></a></li> |
| | + | <li><a href="https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week22"><strong>Week 22</strong></a></li> |
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| - | * '''Team bacterial laccase''': Restriction of ''E. coli'' CueO, ''Xanthomonas campestris'' CopA and the ''B. pumilus'' laccase CotA and the pSB1C3 + RFP plasmid with the same restriction enzymes, ligation of the digested products and transformation in E.coli KRX cells.
| + | </ul> |
| - | ** Since we have less CopA and Tth-laccase DNA, we set an PCR from the remaining PCR approach. For the reaction look [https://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week1 PCR table]
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| | | | |
| - | ===Tuesday May 29th===
| + | </div> |
| - | * '''Team Activity Tests''': Our ordered laccase and ABTS arrived. We couldn't wait to start, so we set up some stocks and created an ultimate plan of how to get to know our laccase better. We found out that natrium acetate buffer (100 mM / pH 5) would give an optimal environment to our enzyme. We decided to check the activity via a photometer. The one we may use here at the Cebitec is a Tecan Microplate reader. Check "protocols" for further information. If oxidized by laccase ABTS can me measured at 420 nm. After some trial and error we found out that a concentration of 0,1 U laccase and 0,1 µl ABTS in each well is perfect for visualizing the process. We added buffer to fill each well to 200 µl.
| + | </div> |
| | + | </html> |
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| - | *'''Team Student Academy:''' ''E. coli'' Mach1 with pMTE cp46 His received from the working group “Fermentation Engineering”, University Bielefeld. Plasmid contains genes for GFP and kanamycine resistance. We plated it and made a liquid culture at 30°C. Result: There was an intense fluorescence. We made a glycerol stock and a plasmid isolation.
| + | <div style="text-align:justify;"> |
| - | | + | ==Week 5 (05/28 - 06/03/12)== |
| - | *'''Team Bacterial Laccases:''' We made colony PCRs on the transformations from the day before. We got product for every transformation approach so we plated the positive colonies on new plates to make plasmid isolation. So hopefully in some days we have the plasmids with the ''E. coli'' laccase, the ''Xanthomonas campestris'' laccase and the ''B. pumilus'' laccase with the inducible t7 promotor and a HIS-tag.
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| - | | + | |
| - | ===Wednesday May 30th===
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| - | *'''Team Bacterial Laccases:''' After plasmid isolation we cut our plasmids with ''Not''I to see if the colony PCR was correct and our laccases are in the backbone. The agarose gel showed that for all of the different plasmids we had at least one plasmid with DNA band on the correct hight in agarose gel.
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| - | | + | |
| - | ===Thursday May 31st=== | + | |
| - | * '''Team bacterial laccase''': We sent the isolated pSB1C3 plasmids with inducible T7 promotor, the different laccase genes from ''Xanthomonas campestris'' CopA, ''B. pumilus'' CotA and ''E.coli'' CueO and HIS-Tag for sequencing.
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| | | | |
| - | ===Friday June 1st===
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| - | ===Saturday June 2nd===
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| - | ===Sunday June 3rd===
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| | | | |
| - | * '''Team Bacterial Laccases''':
| + | {{Team:Bielefeld/Sponsoren}} |
| - | ** PCRs of genomic DNA from ''B. halodurans C-125'' we ordered from DSMZ before. We handled the cells in the same way we did with ''T. thermphilus'' before and soluted the lyophlized cells in water and boiled them before PCR. After PCR we cleaned up the product. However the DNA amount was so low that we had to do the PCRs again.
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