Main Page

Cell and Tissue Culture

• Splitting Cells
• Generating DMSO Stocks for Cell Culture
• Differentiation of 3T3-L1 Cells
• Electroporation of 3T3-L1 Adipocytes
• Fugene Transfection of 293T/COS Cells
• Lipofectamine Mediated Knockdown
• Lipofectamine Plasmid Transfection
• 3T3-L1 Adipocyte Fractionation
• Preparing Cell Lysates
• Glucose Uptake Assay
• Luciferase Assay
• Inositol Labeling and Lipid Extraction

Cloning and Molecular Biology

• Transformation of Bacteria
• Mutagenesis
• PCR Amplification of DNA
• Restriction Enzyme Based Cloning
  • Restriction Enzyme Based Cloning - Ordering Primers
• TOPO Cloning
• Preparing an Agarose Gel
• Cesium Chloride Preparation of DNA

Cell Biology

• Immunofluoresence
  • Colocalization
• FM4-64 Labeling of Yeast Cells

Protein Analysis

• Preparing a SDS-PAGE Gel
• Western Blotting
• Surface Plasmon Resonance - Protein Lipid Interactions

Protein Quantification

• Bradford Assay
• Quantification by Absorbance at 280nm
• Determining Percent Purity
• Using ImageJ to Quantify Bands

Protein Purification

• French Press
• Purification of GST Fusion Proteins
• Affinity Purificatoin of Antibodies (Coupling to Activated CH Sepharose 4B)
• Preparation of DIG Labelled Probes
• Glycogen Synthase Assay
• Triglyceride Assay from Cells and Tissues

Transcriptional Analysis

• Real Time PCR From Cell Culture
• Harvesting RNA from Cells grown in monolayer
• Using Bioconductor To Analyse Beadarray Data
• Using Bioconductor To Analyse Microarray Data

Mouse Protocols

Genotyping

• Ear Tagging and Tail Clipping
• DNA Preparation from Tail Clip
• PCR Analysis of Tail DNA
• Colony PCR

Metabolic Measurements

• Glucose Tolerance Test
• Insulin Tolerance Test
• Measuring Fasting Insulin
• Monitoring Food Intake

Tissue Preparation

• Harvesting Mouse Tissue
• Preparation of Protein Lysates from Mouse Tissues
• Preparation of RNA Samples from Mouse Tissues

Media and Buffers

Yeast Media

• YPD Media and Agar
• Yeast Selective Media and Agar

Buffers

Lysis Buffers

• KRBH Buffer
• RIPA Buffer
• CHAPS Lysis Buffer

SDS-PAGE Buffers

• SDS-PAGE Separating Gel Buffer
• SDS-PAGE Stacking Gel Buffer
• TORC1 Kinase Buffer
• Acrylamide Solution

Other Buffers

• KRBH Buffer
• MTORC1 Kinase Assay
• TORC1 Kinase Buffer

Figure Generation and Data Management

• Generation of Figures in Illustrator
• Using Bioconductor To Analyse Microarray Data