Team:Bielefeld-Germany/Labjournal/week20

From 2012.igem.org

(Difference between revisions)
(→Saturday September 15th)
(→Week 20 (09/10 - 09/16/12))
 
(94 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Bielefeld/Head}}
{{Team:Bielefeld/Head}}
-
<div style="text-align:justify;">
 
-
==Week 20 (09/10 - 09/16/12)==
+
<html>
 +
<style type="text/css">
-
===Monday September 10th===
+
ul {list-style-image:none;}
-
* '''Team Cultivation & Purification:'''
+
#bodyContent{
-
** Cell disruption via high-pressure homogeniser and purification via Ni-NTA column were performed for the following samples: 3L fermentation of ''E.coli'' KRX with laccase from ''B.pumilus'' (09/09), 6L fermentation of ''E.coli'' KRX with laccase from ''E.coli'' (09/07) and ''B.pumilus'' (09/09).
+
    background-color: white;
-
** We made SDS-Pages of purification of laccase from ''B.pumilus''
+
}
-
* '''Team Site Directed Mutagenesis:'''
+
-
**Digestion of the six tvel10-plasmids. Three were unmutated and other three hadn't lost the illegal ''Spe''I-restriction-site, but their second fragment was of a smaller size.
+
-
**plated three additional tvel-t243g-colonies.
+
-
===Tuesday September 11th===
+
</style>
-
* '''Team Shuttle Vector:''' Digest of shuttle shuttle vector with ''Pvu''II and ''Hind''III as control. Agarose gel looks good.
+
-
* '''Fungal Laccases:''' PCR on tvel5 laccase for cloning in shuttle vector. Digest of shuttle vector and digest of tvel35 with ''Aar''I enzyme.
+
-
===Wednesday September 12th===
+
<!-- navigator -->
-
* '''Team Site Directed Mutagenesis:'''
+
<div id="nav" class="tabs">
-
**Plasmid-isolation of the three tvel10-plasmids and digestion with ''Spe''I showed two unmutated plasmids and one with the same wrong restriction-fragments as Monday. There must be a systematical error. ''pfu''-PCR should be done again.
+
<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>
-
===Thursday September 13th===
+
</ul>
-
===Friday September 14th===
+
-
* '''Team Cellulose Binding Domain:'''
+
-
**Isolated three glowing colonies with the [http://partsregistry.org/Part:BBa_K863122 constitutive GFP_His], three with  ClosF 36-38 and Cex 43, 45, 46
+
-
===Saturday September 15th===
+
</div>
-
* '''Team Cellulose Binding Domain:'''
+
</div>
-
**KRX culture of [http://partsregistry.org/Part:BBa_K863103 CBDcex(T7)-GFP_His] seems to have a green glow.
+
</html>
-
**Isolation of four pellets of [http://partsregistry.org/Part:BBa_K863103 CBDcex(T7)-GFP_His] for us and SDU Denmark.
+
-
**[http://partsregistry.org/Part:BBa_K863112 CBDclos(T7)] cut with SpeI+AgeI and deposphorylated.
+
-
**[http://partsregistry.org/Part:BBa_K863120 GFP_Freiburg] (PCR) Gel-clean cut with ''Spe''I and ''Ngo''MIV.
+
-
**Ligated and plated on select-Agar.
+
-
**Restriktionanalysis showed that all GFP_His plasmids are correct, as are the three [http://partsregistry.org/Part:BBa_K863111 CBDclos] and two of the [http://partsregistry.org/Part:BBa_K863101 CBDcex]
+
-
**Collect data to make a protocol for a Cellulose binding assay:
+
-
***Avicel: about 11,4 mg protein (CBD) binds to 1 g Avicel (0,14 mg/12,3 mg)
+
-
***Duration of incubation for CBD to bind to Avicel:  about 30 minutes
+
-
***Washing and Lysis-buffer: 50mM Tris-HCl (pH8.0)
+
-
***If needed: elution with 80% EG or 1/5 Pellet to 4/5 EG (100%) of the overall volume.
+
-
===Sunday September 16th===
+
<div style="text-align:justify;">
-
* '''Team Cellulose Binding Domain:'''
+
-
**Colony-PCR of 15 colonies from the [http://partsregistry.org/Part:BBa_K863113 CBDclos(t7)+GFP_His] transformation-plate (biotaq - Armins recipe) with the result of the positive clones.
+
-
**Prepared the sequencing of GFP_Freiburg 6-8 (1-3)  CBDclos 1-3 (36-38) and CBDcex 1-2 (43, 45)
+
-
**[http://partsregistry.org/Part:BBa_K863113 CBDcex(T7)+GFP_His] isn't glowing anymore, or never was.
+
 +
==Week 20 (09/10 - 09/16/12)==
{{Team:Bielefeld/Sponsoren}}
{{Team:Bielefeld/Sponsoren}}

Latest revision as of 21:31, 25 September 2012


Week 20 (09/10 - 09/16/12)

55px Logo merck.jpg BioCircle.JPG Bielefeld2012 Evonik.jpg Bielefeld2012 Baxter.png Logo knauer.jpg Logo iit.jpg Bielefeld2012 BIEKUBA.jpg Logo biometra.jpg Logo bio-nrw.png Bielefeld2012 Logo ERASynbio.jpg

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.

Contact Us

Universität Bielefeld
Universitätsstraße 25
D-33615 Bielefeld

info(at)igem-bielefeld.de

Navigate our Wiki

Start

Team

Project

Results

Since Regionals

Human Practice

Safety

Judging

Partner

Retrieved from "http://2012.igem.org/Team:Bielefeld-Germany/Labjournal/week20"