Week 1 Lab: Pipetting

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Pippetting

  • Units
    • Moles (mol) measure the absolute amount of a substance
    • Molarity (M) measures the concentration of that substance in a solution
    • Moles (mol): A unit representing particles (atoms, molecules, etc.).
    • Molarity (M): Concentration defined as moles of solute per liter of solution (mol/L).
  • Conversions
    • 1 L = 1000 mL = 1,000,000 μL
    • 1 M = 1000 mM = 1,000,000 μM
  • Pipette sizes
    • P20, P200, P1000 - each fitting up to 20μL, 200μL and 1000μL (1mL)
  • Equipment
    • Pippette
    • Eppendorf Tube
    • PCR tube strip
    • Reagents:
      • dH2O - distilled water (purified)
      • Gel loading dye - used for ???
  • Assays
    • procedure to see if the thing is there or not
    • thing can be a substance, chemical, entity, bacteria, etc.
    • “see” could be measured qualitatively or quantitatively
  • Serial dilutions
    • What is this?
      • It’s a geometric process which downsamples a concentration.
      • This procedure conveys useful information in multiple areas:
        • For measuring population counts using the human eye, you cannot count anything above 10^2, so a 1mL broth which might contain 10^7-10^9 populants can be downsampled to a 1μL broth. There is an innate assumption that the serial dilution process retains a uniform distribution of the original broth.
        • For virology/immunology, you define strength by the last dilution that still works (neutralizes, infects, agglutinates)
        • Dose–response curves - these are log-spaced. The serial dilution process is in a sense a geometric process (reduces by a ratio 1:10 each step, which progressively downscales in logarithmic sense).
      • Serial dilution is how you map an unknown huge concentration into the measurable window of any detector
    • How do you dilute?
      • C1 * V1 = C2 * V2
      • rearrange: V1 = (C2*V2) / C1
      • V_water = V2 - V1
    • How do you do serial dilutions?