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Cell Culture Basics - Transfection
In this lab you will transfect a melanoma cell line using a lipid-based reagent in order to overexpress a gene that is involved in skin cancer.

About This Simulation

In the Cell Culture basics – Transfection simulation you will transfect a melanoma cell line using a lipid-based reagent in order to overexpress a gene that is involved in skin cancer. You will prepare the cells and the reagents and you will use a DNA plasmid carrying the green fluorescent protein as reporter. Will you be able to get fluorescent cells?

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Learning Objectives

  • Understanding the advantages of using lipid-based transfection methods
  • Understanding how to design a transfection experiment:
    • Transfection strategies
    • Use of controls
  • Learning the key steps for transfecting mammalian cells using Lipofectamine 3000:
    • Preparation of cells to achieve highest transfection efficiency (number of passage, percentage of confluence)
    • Preparation of reagents (storing, handling, complexation time, incubation time)
  • Understanding the molecular mechanism of lipid-based transfection
  • Learning how to solve poor transfection efficiency outcomes

Techniques

  • Cell culture
  • Cell transfection
  • Fluorescence microscopy

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