Protocols

• General Reference
• Bioassays
• Cell Culture
• DNA
• RNA
• Protein
• Subcloning
• Transfection
• Expression Library Screening
• Zytoflow

General Reference

• Units
• Formulas
• DNA mobility in gels
• Codon chart
• Amino acid symbols
• Restriction enzymes
• Universal primers
• Listing of M13 (pUC) cloning sites
• Equipment

Bioassays

• NB2-cell proliferation assay
• NK-cell cytotoxicity assay

Cell Culture

• Crystal violet assay
• Decidual cells (modified procedure for establishment of primary cell cultures)
• Passaging cells
• RCHO-1 cells
• Thawing cells (Schreiber's protocol)
• Serum thawing and heat inactivation
• Spongiotrophoblast culture
• Trophoendodermal stem cell cultures
• Trypan blue viability test

DNA

• Dialysis
• Isolation of Genomic DNA (either S.Kwok or H.Mueller)
• PCR
• Phenol / Chloroform / Ethanol - Precipitation
• Sequencing (Sequenase)
• Sequencing (dRhodamine Cycle Sequencing either with GeneAmp 2400 or the DNA Thermal Cycler 480)
• Small Scale Preparation of Plasmid DNA (Miniprep)
• Southern Blot

RNA

• In Situ Hybridization Using DIG-Labeled cRNA Probe

Protein

• Amicon Spiral Cartridge Concentrator
• Amicon Stirred Ultrafiltration Cells
• Alkaline Phosphatase in situ assay (English, Deutsch)
• DEAE - Dextran transfection-COS7 cells
• SEAP-fusion protein binding assay
• SDS-PAGE (Denaturing Discontinuous Gel Electrophoresis)
• Western Blotting
• Dot blot (Western)
• Immunoaffinity chromatography of alkaline phosphatase-tagged proteins

• Immunocytochemistry

  • Bouin's Fixative

  • Acetone Fixative

• Immunoprecipitation
• Protein Assay (Spectrophotometer)
• Protein Isolation / Concentration
• Zymography for Gelatinase-B

Subcloning

• Long Term Storage of Transformed E.coli
• Transformation of competent cells
• Screening of colonies by hybridization

Transfection

• Stable Transfection (Electroporation)

Expression Library Screening

• Titration of Expression Library
• COLT (Cloning of ligand targets)
• Expression Cloning

Zytoflow

• Zytoflow-Analyse von PBL mit SEAP-Fusionsproteinen

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