Protocol
Tools
Plasmids
Cell Stock
Learning Space
Comment
Preparation of LB liquid media and agar plates
Bacterial Glycerol Stock
Plaque Assay
Preparation of Competent Cells
Transformation
MiniPrep of plasmid DNA
MaxiPrep of Plasmid DNA
Preparation of Genomic DNA from Mammalian Cells
Phenol/CIA Purification of DNA
Iso-Propanol Precipitation of DNA
Agarose Gel for DNA and RNA analysis
DNA extraction from Agarose Gel
Preparation of total RNA (Link to RNA Stat-60 Reagent)
Northern Blot Analysis
Real Time RT-PCR for mRNA Quantitation
PCR
Reverse Transcription
for RT-PCR
PCR Screening of Recombinant Colonies
PCR Directed Point Mutation
Restriction Analysis of DNA
DNA Ligation
Preparation of GST fusion protein
SDS-PAGE Electrophoresis
Preparation of GST Fusion Proteins
Preparation of
Biotinylated
Proteins
Immunoblotting (Western Blotting)
Far Western Blot
Antibody stripping and reprobing of Western blot membranes
Immunoprecipitation
ELISA
Reconstitution, propagating and freezing cell lines
Isolation of primary fibroblasts
Trypan Blue Cell Counting
Transfection (LipofectAmine 2000, link PDF)
Transfection of Plasmid DNA into Mammalian Cells (SuperFect)
Transfection of Plasmid DNA into Mammalian Cells (Calcium Phosphate)
Nucleofection
Preparation of Retrovirus
Infection
DNA-Directed RNA Interference
Pulse-chase
Preparation of condition medium (CM) / serum-free medium (SFM)
Preparation of Sucrose Gradients for Cell Fractionation
Immunofluorescent Stain
(adherent cells)
Soft Agar Assay
Zymography of MMP2 and MMP9
Preparation of Collagen coated plate
Collagen Degradation Assay
Invasion Assay 1 (3D Cell Culture On Collagen)
Invasion Assay 2 ( 3D Cell Culture In Collagen)
Invasion Assay 3 (Metrigel)
Immunofluorescent Stain
(tissue sections)
Paraffin-embedded Tissue Block Sectioning &
H&E Stain of Tissue Sections
Labeling Efficiency Measurement by TCA counting
Chase Experiment and Immunoprecipitation with Yeast Cells
Radiation Treatment of Mammalian Cells
Animal anesthesia
Hair removal for surgery
Subcutaneous Injection
(link)
2
Renal capsule grafting
(link)
Tail vein injection for fluorescence imaging
Equipments Instructions
Spectrophotometer
Zeiss Fluorescence Microscope
(Model
Axioplan 2
)
Typhoon Imager
Autoclave Machine
HPLC
Microtome
Polard UV Imager?