Meetings

From 2008.igem.org

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'''Ca-Messung'''<br>
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<li> date for measurement at the AFM (Nitschke)  
<li> date for measurement at the AFM (Nitschke)  
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<li> add wash-steps after addition of DNA-origami
<li> add wash-steps after addition of DNA-origami
<li> cells should keep in Mg2+ for measurement  
<li> cells should keep in Mg2+ for measurement  
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<li> Eventuell könnte man die Messung auch als ELISA durchführen --> immobilisierter AK --> daran bindet das DNA Origami --> 1. die an bestimmte Oligos gekoppelten Fluorophoren können dann detektiert werden --> 2. Man könnte auch Oligos mit Biotin für die DNA Origamis verwenden (sind im Labor vorhanden) und über Streptavidin und ein gekoppeltes Enzym eine Enzymreaktion auslösen
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<li> possibly the measurement can be carried out by ELISA --> immobilized AK to bind origamis; fluorophors fused to the oligos can be detected  '''or'''  use biotin-fused Oligos to get a detection by streptavidin fused enzyme reaction.  
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<li> Kontrolle ob der monoklonale AK an das DNA Origami binden kann (Messung im AFM) --> Daniel anrufen (Simone)
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<li> control: monoclonal AK able to bind DNA-origami (Simone)
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Revision as of 15:52, 29 October 2008


Freiburg2008 small header.gif



Home

The Team

Project Report

Parts

Modeling

Notebook

Safety

CoLABoration

_meetings




Sept. 18th 2008

attendees
Philipp Mappes, Katja Arndt, Kristian Müller, Normann Kilb, Robert Gawlik, Simone Weber, Kathrin Pieper, Sabine Jägle

organisatory

  • Bioss agrees to sponsorship
  • team picture for wiki
  • next meeting: wednesday, Sept. 24th 15.30
  • lab: new box-arrangement (Normann)
  • table for cloning strategy (Kathrin)

Ca-measurement

  • date for measurement at the AFM (Nitschke)
  • optionally: measurement at the Henneke lab --> children's clinic (Sabine)
  • optionally: measurement at Imaging Facility in the medicin clinic --> searching for authorized contact-persons

Wiki

  • carry over the inofficial wiki content to the official one (Robert und Philipp).
  • final report about the whole project (everyone)

part-order

  • ask for the ATG orders (need to send the plasmid again?) (Philipp)
  • obscurities at order BCR-transmembraneregion --> control and correction(Kathrin)
  • primer-order to synthesize signalpeptide by PCR (Kathrin)

control of DNA-Origami binding to cell-surface

  • add wash-steps after addition of DNA-origami
  • cells should keep in Mg2+ for measurement
  • possibly the measurement can be carried out by ELISA --> immobilized AK to bind origamis; fluorophors fused to the oligos can be detected or use biotin-fused Oligos to get a detection by streptavidin fused enzyme reaction.
  • control: monoclonal AK able to bind DNA-origami (Simone)



Sept. 24th. 2008
...


Sept. 29th 2008
...


Oct. 7th 2008'
...


Oct. 17th 2008

attendees:
Philipp Mappes, Katja Arndt, Kristian Müller, Normann Kilb, Robert Gawlik, Michael Kneib

Final report

  • structure equal to a scientific paper containing the parts Abstract, Introduction, Materials and Methods, Results
  • fusion of all sections we worked in: Origami, AFM, FACS, CA2+/Fura measuremt, cell cultures, modeling, ethics
  • quotation of literature through autor and year of publication in the text (Max Muster, 2007), full quotation in references in the end
  • deadline Friday Oct. 24th 2008

official wikipediasite

  • continue the labjournal on the official page only
  • not too much linking: maintenance of the flow of information
  • all embedded files should start with "Freiburg2008"

iGEM parts

  • uploading the parts starting on Monday Oct. 20th 2008



Oct. 21th 2008

attendees:
Moritz Busaker, Philipp Mappes, Kristian Müller, Normann Kilb, Robert Gawlik, Michael Kneib, Kathrin Pieper, Simone Weber


Cloning:
At least - All the constructes are cloned!!!

construcs:

pMA - singalpeptide – scFv-anti-NIP – gggs-Linker – transmembran region - X – pMA

X :

  • bla1(1/2 beta-Lactamase)-
  • bla2 (1/2 beta-Lactamase) –
  • split-linker-C-CFP(Cerutan) –
  • N-CFP –
  • split-linker-C-GFP (split venus)–
  • N-GFP –
  • luciferase 1 (58) –
  • luciferase 2 (57) –



Next steps:
1) We want to test, if the transmembran region really is located in the membrane
Therefore we want to clone a constructe which expresses a YFP on the cytoplasmic side of the receptor.
Construct:
- singalpeptide – scFv-anti-NIP – gggs-Linker – transmembran region - split-linker - bla1 - YFP -

2) We also want to test, if we able to bringe the receptors together by using a NIP coupled peptide.
Therefore we need the peptides from Schamels group -> Norman will ask them.

3) Origami-We want to try if the Origami are able to bring two or more receptors together.
Have to produce new Origami -> Michael and Norman

Next meeting: Oct. 23th 2008

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