Research Methods Flashcards
What makes a good PCR primer?
High GC content G or C at both ends
What does western blotting analyze?
proteins
what type of enzyme is used in southern blotting?
restriction endonuclease
Desolvation
removal/dissociation of the solvent, drying out the sample
Elution
extraction of one material from another via washing can require addition of a buffer to prevent from pH damage from the wash
SDS-PAGE is also known as
gel electrophoresis
SDS-PAGE separates macromolecules by _____
size (and/or charge)
Gas chromatography separates gas molecules by ______
affinity
Mass spectrometry measures the _____ of _________
size
1 molecule
TLC separates molecules by _____
affinity/polarity
stationary phase - polar
mobile phase - nonpolar
Western blotting uses ______ to visualize proteins
antibodies they recognize the antigens on the surface of the proteins
the goal of IR spectroscopy is to determine __________ ______ within a sample
functional groups
IR works by….
hitting with IR beam and measuring the vibration states of the bonds
Diatomic molecules will give ____ _______ in IR because they do not have a ______
no signal dipole
NMR =
nuclear magnetic resonance
Reverse-phase chromatography
same as TLC except mobile phase is polar // stationary nonpolar
column chromatography
seaparates sample in column by polarity
stationary = polar (silica) gel/powder
mobile = nonpolar
Ion-exchange chromatography
separates sample by charge
stationary = charged beads in column
mobile = nonpolar
size-exclusion chromatography
separates sample by size
stationary = polar, porous beads in column
mobile = nonpolar
small molecules end up trapped in pores and elute slower
affinity chromatography
used to purify a molecule of interest
stationary = beads coated with antibody or receptor for target molecule
mobile = nonpolar
gas chromatography (GC)
separates gas compounds
stationary = crushed metal/polymer
mobile = inert gas
HPLC chromatography
essentially column, but more precise
stationary = small column with concentration gradient
mobile = nonpolar
simple distillation
separate 2 liquids with boiling points
below 150C
25C apart
vacuum distillation
for liquid with boiling point above 150C
(lower pressure = lower BP instead of degrading product)
