Protocols.

Preparation of solutions.

• The densities of liquids.
• Salt solutions (part one).
• Salt solutions (part two).
• Single component solution.
• Antibiotics.
• Media, vitamins etc.
• Compound solutions.



Methods from groups of H.Lehrach lab.

Konrad Büssow (Harald Seitz) (buessow@molgen.mpg.de).

• Antibody Detection on high density protein filters.
• Bakterienkultur.
• Elektrokompetente Zellen.
• Expression 96well MTP.
  
• NiNTA microtitre plate purification.
• Non-denat. NiNTA MTP purification.
• Protein purification.
• E.coli mini protein expression.
• Preparation of whole cell. and soluble protein extract.
• Western Blot.




Heinz Himmelbauer (himmelbauer@molgen.mpg.de).

• Amplicon preparation and amplification.
• Embryonic stem cell culture - basic procedures.
• Isolation and handling of mouse embryonic fibroblasts.
• Preparation of genomic DNA from Medaka fish.
• Purification of closed circular DNA by equilibrium centrifugation in CsCl-Ethidium Bromide gradients.




Michal Janitz (janitz@molgen.mpg.de).

• Denatured Salmonsperm DNA.
• Electroporation.
• Oligonucleotide fingerprinting.
• Large Scale PCR.
• Oligofingerprinting protocol.
• Oligo hybridization in Hybridization Bottles using Ready mixes.
• Preparation of 22x 22 cm Agar plates.
• Preparation of electrocompetent Cells.
• Processing of in situ filters.
• Processing PCR filters.
• Radioactive Labeling of a DNA Size Marker.
• Radioaktive "Longprobe"-Hybridization.
• Spotting of PCR-Membranes ("wet").




Aleksey Soldatov (soldatov@molgen.mpg.de).

• 384-well PCR.
• Random primer labeling of polyA+RNA.
• Membrane processing.




Johan Gobom (gobom@molgen.mpg.de).

• alfa-Cyano-4-hydroxycinnamic acid affinity sample preparation. A protocol for MALDI-MS peptide analysis in proteomics.
• Large-gel two-dimensional electrophoresis-matrix assisted laser desorption/ionization-time of flight-mass spectrometry:An analytical challenge for studying complex protein mixtures.




Patricia Ruiz (ruiz@molgen.mpg.de).

• Hybridisation with Nylon-Macroarrays (*.rtf file - save on the disk!).
• Whole-mount in situ hybridization on zebrafish embryos (*.rtf file - save on the disk!).




Silke Sperling (sperling@molgen.mpg.de).

• Complex-Hybridization Protocol for 22x22 filters.
• SSC/SDS based Stripping method.
• Total RNA isolation with TRIzol.
• Hybridisation protocol for 1/6 sized Hybond N filters.
• Evaluierung verschiedener Methoden zur Extraktion von eukaryotischer DNA aus Vollblut.
• Silver staining.




Wilfried Nietfeld (Nietfeld@molgen.mpg.de).

• Super Amine substrate glass microarray processing protocol.
• Array It Microarray glass slide treatment protocols.
• Qualitybackgroundhybridization on glass-microarrays.
• Cy3-/Cy5- labelling of total RNA or mRNA probes.




Marie-Laure Yaspo (yaspo@molgen.mpg.de).

• Post-processing of spotted glass-slides.
• Hybridization of glass-slides.
• Formaldehyde gel for RNA electrophoresis.
• Alkaline gel for cDNA electrophoresis.
• Total RNA isolation from HeLa cell (DNA free).
• Labeled cDNA synthesis and hybridization on nylon membranes.




Eckehard Kuhn (kuhn@molgen.mpg.de).

• Easy plasmid prep.
• TRIzol method for RNA isolation.
• Yeast - glycerol stocks in microtiter plates.
• Yeast - beta-gal assay.
• Yeast - transformation (large-scale).




Pia Aanstad (aanstad@molgen.mpg.de).

• Zebrafish In situ hybridisation protocol.
• Protocol for filter complex hybs.




Georgia Panopoul (panopoul@molgen.mpg.de).

• Protocol for insitus on amphioxus or sea urchin embryos.




Automation group protocols (fritz@molgen.mpg.de).

• cDNA Synthesis.
• Slide Processing.
• cDNA Purification with Microcon YM-30 columns.
• Purification of labelled cDNA with Amersham AutoSeq G50 Columns .
• Oligo Hybridization / Microarray QC.
• Hybridisation procedure with Cy3- / Cy5- labeled cDNA probes.
• RNA Extraktion mit heissem Phenol.