Team:Saarland/Test

From 2014.igem.org

(Difference between revisions)
 
(113 intermediate revisions not shown)
Line 6: Line 6:
<div id="wikicontent-container">
<div id="wikicontent-container">
<div id="wikicontent">  
<div id="wikicontent">  
-
<br>
+
<html>
-
<h1> Labbook </h1>
+
<iframe src="//player.vimeo.com/video/109005558" width="500" height="281" frameborder="0" webkitallowfullscreen mozallowfullscreen allowfullscreen></iframe> <p><a href="http://vimeo.com/109005558">Saarland University iGEM 2014 - Campus Tour</a> </p></html>
-
<br>
+
-
<h2> February</h2>
+
-
*Develop strategy for the first ever biotechnological production of the naked mole rat´s high molecular mass hyaluronic acid and the characterisation of its inhibitory effects on carcinogenesis.
+
-
*Recruiting committed team members.
+
-
 
+
-
<h2> March </h2>
+
-
*Looking for sponsors.
+
-
*Team registration.
+
-
 
+
-
<h2>April</h2>
+
-
*Looking for sponsors
+
-
*Design of a team logo
+
-
 
+
-
<h2>May </h2>
+
-
*Looking for sponsors
+
-
*Conception and design of the team wiki
+
-
 
+
-
*Waiting for completion of <i>has</i>2 gene synthesis.
+
-
 
+
-
<h3>Highlight at Week 4</h3>
+
-
*Meet up of German iGEM teams in Munich. Inspiring Weekend. We enjoyed the short trip.
+
-
 
+
-
<h2>June</h2>
+
-
<h3>1. Week</h3>
+
-
*Finally, the synthesised <i>has</i>2 gene has arrived!
+
-
 
+
-
*Transformation of electro competent <i>E. coli</i> cells with Eurofins standard delivery plasmid pexK4 containing the synthesised <i>has</i>2 insert DNA.
+
-
 
+
-
*Plasmid preparation and control digestion to confirm the presence of <i>has</i>2 insert DNA.
+
-
 
+
-
 
+
-
 
+
-
<h3>2. Week</h3>
+
-
*First attempt to insert <i>has</i>2 DNA into the multiple cloning site (MCS) of pSMF2.1 plasmid construct. This shuttle plasmid allows on the one hand the amplification of plasmid DNA in high copy numbers in <i>E. coli</i> cells. On the other hand pSMF2.1 is optimised for expression of recombinant proteins in our favourite chassis <i>B. megaterium. Has</i>2 insert DNA was absent in transformants.
+
-
 
+
-
<h3>3. Week</h3>
+
-
*Repetition of cloning experiments (week 2) with new T4 ligase provided many <i>E. coli </i>transformants with pSMF2.1 plasmid construct containing <i>has</i>2 insert DNA.
+
-
 
+
-
<h3>4. Week</h3>
+
-
*Extraction of genomic DNA from <i>B. megaterium</i> was performed.
+
-
 
+
-
*Optimal PCR conditions for the amplification of genomic DNA from <i>B. megaterium </i>were tested. First gene (<i>UDP-glc</i>DH) involved in synthesis of precursor molecules and probably contributing to a high yield production of high molecular mass hyaluronic acid was successfully amplified.
+
-
 
+
-
*First attempt for downstream insertion of <i>UDP-glc</i>DH DNA into the MCS of pSMF2.1 plasmid construct that already contained the <i>has</i>2 insert DNA was successful.  
+
-
 
+
-
<h2>July</h2>
+
-
<h3>1. Week</h3>
+
-
*Open day at Saarland University. The iGEM team was proud to present its project to the general public.
+
-
 
+
-
*Plasmid preparation and control digestion to confirm the simultaneous presence of <i>has</i>2 and <i>UDP-glc</i>DH insert DNA in the pSMF2.1 plasmid construct.
+
-
 
+
-
*Preparation of fresh protoplasts for transformation of <i>B. megaterium</i> strain MS941 was successful. Competence is outstanding.
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<h3>2. Week</h3>
+
-
*Transformations of <i>B. megaterium</i> protoplasts with different pSMF2.1 plasmid constructs were successful. Screening process for the identification of <i>B. megaterium</i> transformants with correct plasmid constructs was time consuming.
+
-
 
+
-
*<i>B. megaterium</i> chassis for the first ever biotechnological production of high molecular mass hyaluronic acid is ready!
+
-
 
+
-
<h3>3. Week</h3>
+
-
*Cultivation of <i>B. megaterium</i> and induction of recombinant protein production were performed. Samples were prepared after an appropriate cultivation time to check and characterise recombinant protein expression levels of Has2 and UDP-GlcDH.
+
-
 
+
-
*These samples were also used for viscosity measurement since the outstanding water binding capacity of high molecular mass hyaluronic acid should increase the viscosity of the cultivation medium.
+
-
 
+
-
<h3>4. Week</h3>
+
-
*Illegal restriction sites within <i>has</i>2 and <i>UDP-glc</i>DH were changed via QC-PCR. Resulting biobrick DNAs were inserted into the standard biobrick plasmid construct pSB1C3.
+
-
 
+
-
*Measurement of viscosity was performed. There was no difference in viscosity of the culture medium and consequently no high molecular mass hyaluronic acid production detectable in the first attempt.
+
-
 
+
-
<h2>August</h2>
+
-
<h3>1. Week</h3>
+
-
*SDS-PAGE
+
-
 
+
-
*Power failure at whole Saarland University during weekend. Reorder of heat sensitive enzymes including polymerases, restriction enzymes and ligase.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project
+
-
 
+
-
<h3>2. Week</h3>
+
-
*Functional enzymes arrived.
+
-
 
+
-
*Repetition of all <i>B. megaterium</i> transformations during timeout since corresponding glycerol stocks were damaged.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project
+
-
 
+
-
 
+
-
<h3>3. Week</h3>
+
-
*Generation of <i>has2-gfp</i> fusion gene via SOE-PCR for quick evaluation of Has2 expression level and localisation in <i>B. megaterium</i> was performed. Insertion of <i>has2-gfp</i> fusion gene into MCS of pSMF2.1 plasmid construct was successful. A lot of positive <i>E. coli</i> transformants!
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project
+
-
 
+
-
 
+
-
<h3>4. Week</h3>
+
-
*Plasmid preparation and transformation of fresh prepared <i>B. megaterium</i> protoplasts with pSMF2.1 plasmid construct containing the <i>has2-gfp</i> fusion gene was successful.
+
-
 
+
-
*Expression of Has-GFP fusion protein and samples for localisation studies via fluorescence microscopy were prepared.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project
+
-
 
+
-
 
+
-
<h2>September</h2>
+
-
<h3>1. Week</h3>
+
-
*Meet and greet with our sponsors. Thank you very much for your great support!
+
-
 
+
-
*First attempt to confirm Has-GFP expression via epi fluorescence microscopy was successful. <i>B. megaterium</i> cells glew beautifully green.
+
-
 
+
-
*Additional genes involved in synthesis of precursor molecules were successfully amplified from genomic DNA of <i>B. megaterium</i> and inserted into the standard biobrick plasmid construct pSB1C3.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
* Evaluation of results and creation of figures for our team wiki.
+
-
 
+
-
 
+
-
<h3>2. Week</h3>
+
-
*Expression of Has-GFP fusion protein was repeated and samples for localisation studies via laser scanning microscopy were prepared. No fluorescence signal was detectable after cultivation under the same conditions.
+
-
 
+
-
*Illegal restriction sites within the genes for pathway engineering were changed via QC-PCR.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
*Evaluation of results and creation of figures for our team wiki.
+
-
 
+
-
 
+
-
 
+
-
<h3>3. Week</h3>
+
-
*Expression of Has-GFP fusion protein was tested under different cultivation conditions. No significant fluorescence signal. Possible misfolding of Has-GFP protein
+
-
 
+
-
*Sequencing of biobricks was prepared.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
*Evaluation of results and creation of figures for our team wiki.
+
-
 
+
-
 
+
-
<h3>4. Week</h3>
+
-
*Waiting for sequencing results.
+
-
 
+
-
*Human Practices: Design and development of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
*Evaluation of results and creation of figures for our team wiki.
+
-
 
+
-
 
+
-
<h2>October</h2>
+
-
<h3>1. Week</h3>
+
-
*Biobrick sequencing was finished. Biobricks are ready to ship to the iGEM Headquarter.
+
-
 
+
-
*Human Practices: Beta test of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
*Evaluation of results and creation of figures for our team wiki.
+
-
 
+
-
<h3>2. Week</h3>
+
-
*Last modifications on team wiki!
+
-
 
+
-
*Human Practices: Last modifications of the roleplaying game for the ethical and social implications of our project.
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
</div>
</div>
</div>
</div>
{{Team:Saarland/footer}}
{{Team:Saarland/footer}}

Latest revision as of 12:39, 17 October 2014


Retrieved from "http://2014.igem.org/Team:Saarland/Test"