Stephens Lab Protocols

Core Protocols

• Making competent E. coli cells

• Fugene transfection (HeLa cells)

• Lipofectamine 2000 transfection (A549, McA7777 cells)

• High efficiency transfection of Vero cells

• Immunoprecipitation

• Protein kinase assays

• Molecular Biology methods

• Purification of Sar1

• Silver staining polyacrylamide gels

• Yeast media recipes

• Plasmid isolation from yeast

• Preparation of yeast protein extracts

• Colony lift filter assays for two hybrid screening

Cell Imaging Protocols

• Volumes and areas of tissue culture dishes and multiwell plates

• Fibronectin coating

• Making Mowiol for mounting coverslips

• Immunofluorescence (basic protocols)

• Incorporation of BrdU to measure DNA replication

• Optiprep methods (link to Axis Shield website)