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

Protein Purification

• French Press
• Purification of GST Fusion Proteins
• Affinity Purificatoin of Antibodies (Coupling to Activated CH Sepharose 4B)
• Preparation of DIG Labelled Probes

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