Green Fluorescent Protein Lab Essay

1704 WordsMar 16, 20067 Pages
Title: Purification of Green Fluorescent Protein Introduction: Transformation is used to introduce a gene coding for a foreign protein into bacteria. Hydrophobic Interaction Chromatography (HIC) is used to purify the foreign protein. Protein gel electrophoresis is used to check and analyze the pure protein. Research scientists use Green Fluorescent Protein (GFP) as a master or tag to learn about the biology of individual cells and multicultural organisms. This lab introduces a rapid method to purify recombinant GFP using HIC. Once the protein is purified, it may be analyzed using polysaccharide gel electrophoresis (PAGE). Purpose: To illustrate the process of transformation and perform it using Green Fluorescent Protein.…show more content…
Invert. 16. Microcentrifuge for 1 minute. Use a micropipette to transfer the supernatant containing the recombinant GFP to a new 1.5mL Eppendorf tube. 17. Observe the GFP under ultraviolet light. Data: (Laboratory Procedure for pGREEN) 1./2./3. LB-plasmid: LB+plasmid: LB/Amp+plasmid: LB/Amp-plasmid Prediction: average growth average growth growth no growth Reason: control control it has LB/Amp and plasmid no plasmid Result: lawn lawn 17 colonies no growth 4. LB+plasmid (Pos Control) lawn LB-plasmid (Pos Control) lawn LB/Amp+plasmid (Exper) 17 colonies LB/Amp-plasmid (Neg Control) no growth 5. a. LB+plasmid and LB-plasmid: Both of these plates had a lawn of bacteria. This proves that the bacteria are capable of growing on the agar and that there was nothing preventing growth beside the ampicillin. b. LB/Amp-plasmid and LB-plasmid: The LB/Amp-plasmid had no growth compared to the LB-plasmid which had a lawn. This proves that transformation cannot take place without the plasmid, or DNA. c. LB/Amp+plasmid and LB/Amp-plasmid: The LB/Amp-plasmid had no growth, but the LB/Amp+plasmid had growth. This shows that the bacteria was transformed and
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