Bio Lab

• Personal Protective gear required

  • Goggles
  • Gloves
• Lab coats are available
• Qualitative comparison


  Interpretation-based, descriptive
• Quantitative comparison


  Measurable, defined
• C1V1=C2V2


  1. C1 = stock concentration

  2. C2 = final concentration you want

  3. V1 = unknown volume you're trying to figure out

  4. V2 = the desired final volume you're working with
• Gel electrophoresis


  • Bands are separated by size
  • Need a reference ladder with bands of known size
• Ethidium bromide


  Binds to DNA by intercalating between the bases and fluoresces under UV light
• Bradford assay


  Colorimetric assay used to quantify the protein concentration in a sample
• Glucose oxidase assay


  1. Glycogen levels in tissue can be measured indirectly by measuring the amount of glucose liberated from glycogen

  2. Enzyme digests glycogen into glucose monomers

  3. Glucose is quantified with a colorimetric assay

  
• Cell membranes


  • Selectively permeable
  • Rate of membrane penetrance depends on molecular size, lipid solubility, degree of ionization
  • Molecules that don't fit these requirements can still enter the cell through channels or receptors
• Osmosis


  Water moves across the membrane so that the relative concentration of impermeable solutes is equal on both sides
• Tonicity


  The change in volume of cells in response to a solution
• Isotonic solution of red blood cells is the cloudiest

  
• Measuring succinate dehydrogenase (SDH) activity

  1. SDH catalyzes the reaction: Succinate + FAD ↔ fumarate + FADH2

  2. DCIP serves as an electron acceptor that accepts the electrons that SDH removes from succinate

  3. Oxidized DCIP is dark blue, reduced DCIP is colorless
• Malonate


  Competitive inhibitor of SDH
• You will not need to perform any statistical analysis, but may need to answer a multiple choice or short answer question
• Standard deviation (σ)


  Measure of variation or dispersion between values in a set of data
• n


  Number of data points from each group (how many replicates you did)
• P value


  • Probability that sample differences are due to chance
  • Low p value means the difference is most likely not due to chance
  • P=0.05 is usually the cut off for considering something statistically significant
• ANOVA


  Analysis of variance is a statistical test for detecting differences in group means when there is one parametric dependent variable and one or more independent variables
• Photosynthesis


  1. Light reactions

  2. Calvin Cycle (dark reactions)
• Hydrogen carbonate indicator solution

  Qualitative measurement sensitive to changes in pH caused by CO2 from the air dissolving in water to form carbonic acid

  
• Photosynthesis


  Reduces CO2, increases pH, shifts towards purple
• Respiration


  Produces CO2, decreases pH, shifts towards yellow
• Bacterial transformation


  Bacteria take up exogenous (foreign) DNA and produce the genetic products (proteins) encoded by that DNA
• Bacterial genome


  • Haploid and generally contained on a single circular chromosome
  • Some bacteria also possess additional pieces of circular DNA called plasmids
• Plasmid


  Contains fewer genes than the chromosome but can offer the cell unique characteristics
• Selectable marker


  Provides a way of knowing which cells took up the plasmid
• PCR


  1. Denaturation

  2. Annealing

  3. Extension
• DNA Polymerases


  • Possess 5'→ 3' polymerase activity, which is the incorporation of nucleotides to extend primers at their 3' ends in the 5' to 3' direction