300662 SE Seminar zu Übung III A (2009S)

Ort: BZB/SR 1, 6.Stock, Dr. Bohrgasse 9, 1030 Wien.

Vorbespr: 5.3.09, 17.00 Uhr

Teilnahme verpflichtend, Themenvergabe

Informationen siehe auch Studentenseite des Zentrums für Molekulare Biologie: http://www.students.mfpl.ac.at/

Termine des Seminars (Beginn jeweils 9 Uhr 15, Seminarraum 1, Dr. Bohrgasse 9/6):
18. 5. : Bachmayr-Heyda, Vejdovszky, Zappe
19. 5.: Reinthaler, Halimi, Lippert
22. 5..: Steinkellner, Finkes, Stöger
25. 5.: Hausmann, Anderluh, Gselmann
26. 5.: Paur, Bauer, Fluch
27. 5.: Ramesmayer, Pingger, Pichler
28. 5.: Krenbeck, Kalser, Rohrer

Ort: BZB/SR 3, Dr. Bohr-Gasse 9, 1030 Wien.
Teilnahme verpflichtend, Themenvergabe
Informationen siehe auch Studentenseite des Zentrums für Molekulare Biologie: http://www.students.mfpl.ac.at/

VORBESPRECHUNG: 5. März, 17.00 Uhr, SR1 6. Stock

Vorläufige Anmeldungen vom 5.März mit Seminarthemenliste:

1) Describe the E.coli transformation method(s) and plasmid vectors (such as pUC18/19, pBluescript, pGEM-Teasy or others) by which foreign DNA can be transfered into E.coli.
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2) Describe in detail the isolation of the plasmid DNA from E.coli and explain the theoretical basics behind it (use as basis a method provided in the protocol).

3) Restriction enzymes and DNA modification enzymes (ligase, DNA polymerase, Klenov, SAP etc.). Theoretical background. Their features and use in DNA analysis and modification (restriction analysis, ligation of DNA fragments etc.) (2 students).
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4) Theoretical background of PCR and applications in recombinant DNA techniques (!!!RT-PCR will be a separate topic!!!!).
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5) Describe specification of commonly used E. coli (DH5alpha, XL1-blue, HB101) and yeast strains (such as PJ69-4A, L40, H7Fc) Describe some basic yeast shuttle vectors (pGAD424, pBDcam, pYES), methods of yeast transformation and selection of transformants. Yeast two hybrid assay and Yeast two hybrid screens: theory, applications and plasmid rescue from yeast (group of 3 students).
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6) Describe the methods how to introduce foreign DNA into plants, to receive stable transformants. Describe vectors (e.g. pGreen, pBI101) for plant stable transformation and example of application (group of 2 students).
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7) Main features of vectors for plant transient transformations. Methods of transient plant cells transformation.
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8) Describe the methods to analyse foreign (transgenic) protein expression in plants (group of 2 students):
------------Western blotting, tagging of proteins: different tags, detection.
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9) Describe methods for gene expression analysis (on the RNA level) in plant(eukaryotic) cells (RT-PCR, cDNA library) (!Northern is a separate topic #12)
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10) Microarrays: theory, examples (DNA and oligo chips), CHIP on chip analysis, examples group of 2 students
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11) Southern Blot, Northern Blot and their applications in molecular biology
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12) RNA and DNA in situ hybridisation methods, examples
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13) Describe methods of imaging analysis in eukaryotic (plant) cells, examples.
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Literature:
IMP-library (recommended)
Molecular Cloning: A Laboratory Manual (Library Binding)
by Joseph Sambrook (Author), T. Maniatis (Author) "THIS PRINT EDITION OF MOLECULAR CLONING is associated with a Web Site www.MolecularCloning.com
that is evolving into an on-line laboratory manual... Molecular Cloning: A Laboratory Manual (3-Volume Set), Joseph Sambrook, David W. Russell (original by Tom Maniatis)
Very simplistic but helpful: Instant Notes in : Molecular Biology, Genetics, Plant Biology (BIOS Scientific Publishers)
IMP-library (recommended),
WWW Diverse web sites e.g.:
http://www.yeastgenome.org/VL-yeast.html#buds
http://www.fermentas.com/techinfo/appendix/index.html
http://www.neb.com/nebecomm/tech_reference/default.asp
http://www.arabidopsis.org/servlets/Search?action=new_search&type=protocol
ftp://ftp.arabidopsis.org/home/tair/Protocols/Mundy_Protocols.pdf
wikipedia

Ort: BZB/SR 3, Dr. Bohr-Gasse 9, 1030 Wien.
Teilnahme verpflichtend, Themenvergabe
Informationen siehe auch Studentenseite des Zentrums für Molekulare Biologie: http://www.students.mfpl.ac.at/