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Gene Expression

In this lab you will take samples of lean and obese pigs and analyze the levels of gene expression by PCR, RT-PCR, NGS and qPCR experiments.

About This Simulation

To study obesity you will collect a fat sample from lean and obese pigs and extract the RNA using the phenol-chlorophorm method. You will isolate and purify the mRNA and utilize RT-PCR techinque to synthesize cDNA. After that you will modify the cDNA sample with adaptors to be able to use the Next Generation Sequencing (NGS) technique.

You will perform a NGS experiment, learning all the important steps in this high-tech molecular biology procedure with inmersive 3D animations that show what is happening at the molecular level inside the NGS machine. You will also learn how to analyze the data from the experiments, understanding how mRNA relate to the gene expression levels and how this can affect obesity.

After identifying a candidate gene involved in obesity a quantitative PCR (qPCR) experiment needs to be performe to validate your results. You will prepare the qPCR reaction and obtain the melting and standard curves. Would your analysis confirm your previous results? Would you find a gene responsible for obesity?

Screenshots

Collaborators

Prof. Susanna Cirera Salicio

Prof. Susanna Cirera Salicio

Institute for Department of Basic Animal and Veterinary Sciences

Faculty of Health and Medical Sciences, University of Copenhagen

Learning Objectives

  • Understanding the central dogma of molecular biology
  • Knowing the principles of RNA extraction and mRNA purification
  • Learning about the molecular biology techniques: PCR, RT-PCR, qPCR and Next Generation Sequencing
  • Understanding that cell functions are regulated by gene expression

Techniques

  • RNA isolation
  • cDNA synthesis
  • Next generation sequencing
  • Gene expression
  • qPCR

Collaborator

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