Biochemistry Chapter 5: Analytic Techniques Flashcards
In gel electrophoresis, molecules move from
Cathode (-) to anode (+)
True or false: PCR can be used to quantify gene expression
False. Quantifying gene expression must be performed with other lab techniques, such as qPCR/RT-PCR.
what does PCR require?
Forward and reverse primers
Taq polymerase
A magnesium containing buffer
True or false: retention factor can only be between 0 - 1
True. Its Rf value will thus be quite high. Rf, or the retention factor, is equal to the distance a given solute has migrated up the stationary phase, divided by the maximum distance traveled by the mobile phase, or solvent. This means that Rf can range from 0 to 1; lower Rf values (closer to 0) are associated with more polar compounds, whereas higher values (closer to 1) are associated with more nonpolar compounds.
Rf formula
Beer lambert law
where A is absorptivity, ε is the molar attenuation coefficient, and c is the concentration. Essentially, it indicates that higher levels of absorbance mean that more light is absorbed, which means that more of a substance is present.
technique that assesses the concentration of protein by measuring the extent to which the unlabeled proteins in the sample compete with radioactively labeled antigens for antibody binding sites
Radioimmunoassay
technique in which antibodies bind antigens, and then the amount of bound antibody is determined by reactions that produce a visible signal, like color or fluorescence
ELISA
Rule of thumb for correlation between variables
Fractional distillation is used to separate what kind of compounds?
Compounds with very close boiling points
When to use thin layer chromatography vs gel electrophoresis
TLC = for organic (polar/non-polar)
Gel Electrophoresis = for charged species like DNA
what direction do the proteins migrate in SDS-PAGE?
Towards the positive anode (SDS-PAGE has a current running through it). Like an electrolytic cell
why is albumin often used in biochemical and enzymatic experiments?
In biochemical and enzymatic experiments, albumin (such as BSA) is commonly used to prevent enzyme adsorption to surfaces, stabilize proteins, and maintain reaction homogeneity for accurate kinetic measurements.
