Week 10: Advanced Imaging and Measurment Technology

What happened to my brain?
question question
The Only Question -RCBeck

Final Project Homework

For your final project:

  1. Please identify at least one (ideally many) aspect(s) of your project that you will measure. It could be the mass or sequence of a protein, the presence, absence, or quantity of a biomarker, etc.

  • Aim One: Measurment Data
    The first aim of this project transforms a strain of Synechococcus elongatus 7942 (PCC7942) into the sucrose secreting, model organism S. elongatus cscB+. The original PCC7942 colony will need to be grown to an optical density (OD) 750 of ≥1 before transformation may take place. A spectrophotometer will be used to obtain this measurement.
    - First growth media needs to be prepared, which will require measuring agar, BG11 medium, AS Trace Elements, water, pH, and temperature of growth media. Plates are poured, streaked and incubated. Colonies are then selected and transfereed to new grow media.
    - Colonies are incubated until an OD of 750 is reached, then natural transformation may take place.
    - Transformed samples are then recultured with spectinomycin treated plates, which selects for only transformed model organisms. chart1 chart1

  • Aim Two: Measurment Data
    Transformation of second organsim, E. Coli is the another two stage transformation. The first stage transforms the bacteria from its native state into 3-hydroxypropionic acid (3-HP) producing Escherichia coli. The next stage of transformation will modify 3-HP to express chromo-proteins which will be co-cultured with the S. elongatus GC strain.
    chart2 chart2

  • Aim Three: Measurment Data

  1. Please describe all of the elements you would like to measure, and furthermore describe how you will perform these measurements. What are the technologies you will use (e.g., gel electrophoresis, DNA sequencing, mass spectrometry, etc.)? Describe in detail.