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Contents

  • 1 Cell and Tissue Culture
  • 2 Cloning and Molecular Biology
  • 3 Protein Analysis
    • 3.1 Protein Quantification
    • 3.2 Protein Purification
  • 4 Transcriptional Analysis
  • 5 Mouse Protocols
    • 5.1 Genotyping

Cell and Tissue Culture

  • Splitting Cells
  • Differentiation of 3T3-L1 Cells
  • Electroporation of 3T3-L1 Adipocytes
  • Immunofluoresence
  • 3T3-L1 Adipocyte Fractionation
  • Preparing Cell Lysates
  • Harvesting RNA from Cells grown in monolayer

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

Protein Analysis

  • Western Blotting
  • Surface Plasmon Resonance - Protein Lipid Interactions

Protein Quantification

  • Bradford Assay
  • Quantification by Absorbance at 280nm

Protein Purification

  • French Press
  • Purification of GST Fusion Proteins
  • Determining Percent Purity

Transcriptional Analysis

  • Real Time PCR From Cell Culture
  • Harvesting RNA from Cells grown in monolayer

Mouse Protocols

Genotyping

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