Biotechnology

Part 1: pGLO Bacterial Transformation and DNA Extraction          June 27th, 2017
Transformation: Uptake and expression of a gene from one organism to another
Creates a transgenic organism with recombinant DNA
Cutting and pasting DNA from source to new cells
  • Cutting: Restriction enzymes
  • Pasting: Ligases

In this experiment
Recipient: E. Coli bacterial cells
Vector: pGLO plasmid
  • Genes of interest:
    • GFP gene= green fluorescent protein
    • bla gene= resistant to ampicillin
    • araC gene= allows GFP to be expressed
Standardized variables: LB infused agar




Conclusions: The +DNA LB/ Amp was able to successfully grow colonies, but notice that they don’t glow. The treatment group that was +DNA LB/ AMP/ ARA was able to use the gene for fluorescence with the presence of arabinose sugar.

Part 2: Polymerase Chain Reaction (PCR)          June 29th, 2017
Based on DNA replication
Used in forensic science, detect mutations, paternity testing


Process details:
  1. Denature DNA to break it apart into two strands (95 C)
  2. Anneal primers to bind to target
  3. Extend primers to taq polymerase enzyme

Part 3: Gel Electrophoresis                      July 6th, 2017
Used to identify gene or organism, synthesize new DNA, analyze gene expression, measure DNA
Method of sorting macromolecules (especially proteins and nucleic acids) based on their size and charge

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Without UV Light        With UV Light

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