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| | {{Team:Bielefeld/Head}} | | {{Team:Bielefeld/Head}} |
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| - | <div style="text-align:justify;">
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| - | ==Week 14 (07/30 - 08/05/12)==
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| - | ===Monday July 30th=== | + | <html> |
| - | * '''Team Site Directed Mutagenesis:''' Preparations for plasmid-isolations of ''B.pumi''_2883_IV (I-IV); ''X.camp''_3633 (I-IV); ''X.camp.''_2247_2 (I-IV) and ''B.pumi''_2883_III (I-IV)
| + | <style type="text/css"> |
| - | * '''Team Cellulose Binding Domain:''' Redesigned the primer-sequences another time giving the CBDs a few AS more in than the Protein-BLAST said. CBDcex: 4 AS N-terminal, 2 C-terminal. CBDclos: 2 N-terminal (starting with a natural ATG) and 2 AS C-terminal.
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| - | **Prepared BBa_392014 for Sequencing
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| - | * '''Team Bacterial Laccases:''' Restriction of Tv5 laccase PCR product with Prefix and Suffix ends for cloning in pSB1C3 and restriction of laccase with the overhangs for cloning in shuttle vector. And we did the PCR on Tv35 with both primer pairs again. This time we lowered the annealing temperatures and got products with both primer pairs.
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| | | | |
| - | ===Tuesday July 31st===
| + | ul {list-style-image:none;} |
| - | * '''Team Site Directed Mutagenesis:''' Picked four colonies per dish ''T.vers.''_243 & ''T.vers.''_1161 and plated them for plasmid-isolation.
| + | #bodyContent{ |
| - | **Plasmid-isolation of ''B.pumi'' III(III)+IV(IV) with SDM-Mix 2317 ''X.camp'
| + | background-color: white; |
| - | **Test-restriction 700ng in 10 µL with ''Pst''I
| + | } |
| - | :Results of cutting if mutation failed:
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| - | :The ''B.pumilus'' site directed mutation at 2317 is not a illegal restriction site, so it can not be checked via restriction, but only through sequencing.
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| - | :*Picked a few more Colonies of X.camp for plasmid-isolation
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| - | * '''Team Bacterial Laccases:'''
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| - | :* Ligation of Tv5 laccase with pSB1C3 backbone and purification of Tv35 laccase PCR products.
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| - | :* We designed primers with the sequences of different promoters from the [Enderson promotor family]. We chose the sequences of the parts [http://partsregistry.org/wiki/index.php/Part:BBa_J23103 BBa_J23103], [http://partsregistry.org/wiki/index.php/Part:BBa_J23103 BBa_J23110] and [http://partsregistry.org/wiki/index.php/Part:BBa_J23103 BBa_J23117].
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| | | | |
| - | ===Wednesday August 1st===
| + | </style> |
| - | * '''Team Site Directed Mutagenesis:''' Plasmid isolation of ''T.vers''_243 (I-IV) & ''T.v.''_1161 (I-IV) ''X.camp''_2K&3K (V-VI)
| + | |
| - | **Test-restriction with ''Pst''I / ''Spe''I & Anaylsis via Gelelectrophoresis show bands as they should for ''T.vers''_243 II+III; and ''X.camp.''_2K_2 V
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| - | **Made ''X.camp.''_2K_2 V and the final ''B.pumilus''plasmids ready for sequencing
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| | | | |
| - | ===Thursday August 2nd=== | + | <!-- navigator --> |
| - | * '''Team Wiki:''' This morning we met for taking a new picture of our group and individual pictures of everyone. Check out our beautiful team members [https://2012.igem.org/Team:Bielefeld-Germany/Students here].
| + | <div id="nav" class="tabs"> |
| - | * '''Team Site Directed Mutagenesis:''' SDM PCR of ''T.vers''243 II+III with 1161 Primer mix and ''X.camp.''_2K_2 V with 3633
| + | <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> |
| | | | |
| - | ===Friday August 3rd===
| + | </ul> |
| - | * '''Team Site Directed Mutagenesis:''' Transformed ''T.vers''_243_III_1161 and ''X.camp.''_2K_2 V_3633 in XL1 Blue and plated it on Ampicillin- and CM-selection-agar.
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| - | * '''Team Cellulose Binding Domain:''' Transformed the plasmids p714_cex and 570_clos we got from the fermantationgroup in KRX an plated them on Kanamycin-selectionagar.
| + | |
| | | | |
| - | ===Saturday August 4th===
| + | </div> |
| - | * '''Team Site Directed Mutagenesis:''' Plated four isolated colonies from ''T.vers''_243_III_1161 and ''X.camp.''_2K_2 V_3633 per petri dish on seperated selection-agar-dishes
| + | </div> |
| | + | </html> |
| | | | |
| - | * '''Team Cellulose Binding Domain:''' Since there were only a few colonies on the p570 selection-agar-dish, I picked one, resuspended it in 100 µL destilled water and plated it on an new Kanamycin-selection-agar-dish
| + | <div style="text-align:justify;"> |
| - | **plasmid-isolation of p714
| + | ==Week 14 (07/30 - 08/05/12)== |
| - | | + | |
| - | ===Sunday August 5th=== | + | |
| - | * '''Team Site Directed Mutagenesis:''' Plasmidisolations of ''X.camp.''2K_2_3633 (I-IV) and T.vers_243_III_1161 (I-IV)
| + | |
| - | **Test-Restriktion of ''X.camp.''2K_2_3633 (I-IV) and ''T.vers''_243_III_1161 (I-IV) X.camp III + IV good; T.vers, no positiv, since the 1150 band should at least rise to 1550
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| - | **Picked two additional colonies of ''e.coli''_2307 and plated them on CM
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| - | **Prepared ''X.camp.''2K_2_3633 III + IV for sequencing
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| - | * '''Team Cellulose Binding Domain:''' Transformation of BBa_I13522 + pSB1A2 in KRX and plating it on AMP-Selection-Agar
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| - | **Plasmidisolations of p570
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| - | **PCR of CBDcex (417 bp) and CBDclos (369 bp)
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| - | ***Test-Gelelectrophoresis: Got the bands that should be there!
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| - | ***PCR Clean up (no Gelelectrophoresis)
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| - | **Restriktion of PCR-products and pSB1C3_RFP with XbaI and PstI.
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| | {{Team:Bielefeld/Sponsoren}} | | {{Team:Bielefeld/Sponsoren}} |
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