Chapter 5: Molecular Tools for Studying Genes Flashcards
(34 cards)
sodium dodecyl sulfate (SDS)
denatures proteins and adds negative charge to proteins so they’ll migrate toward the anode
used to determine major landmarks within a strand or segment of DNA: a physical map of locations of restriction sites within piece of DNA
restriction mapping
restriction mapping
used to determine major landmarks within a strand or segment of DNA: a physical map of locations of restriction sites within piece of DNA
aka size exclusion; large proteins move faster b/c smaller ones get stuck in the matrix
gel filtration chromatography
a technique used to separate/purify fragments/strannds of nucleic acids
always includes known DNA ladder
gel electrophoresis
gel electrophoresis
a technique used to separate/purify fragments/strannds of nucleic acids
always includes known DNA ladder
hybridization
the ability of a strand of DNA/RNA to form a double helix with another strand of DNA/RNA
used to separate/purify polypeptides
polyacrylamide gel electrophoresis (PAGE)
invented by Edward Southern; transferred DNA from gel to thin membrane of microcellulose
gel is run and placed in “sandwich” with membrane and exposed to current to transfer DNA. Membrane/filter is then washed, blocked, and exposed to highly specific probe
southern blotting
a powerful tool used for forensics, paternity testing, medical screening, or phylogeny.
A HaeIII restriction cut of a specific region of DNA followed by Southern blot
fingerprinting
placing (loading) DNA/RNA into agarose gel and subjecting to current, causing DNA to migrate toward positive pole
DNA electrophoresis
column chromatography
method for fractionating nucleic acids or proteins using stationary phage from cellular lysate (i.e. nucleic acids in solution)
tracers, labels, probes
highly sensitive techniques used to detect a gene, nucleic acid, or protein of interest
DNA electrophoresis
placing (loading) DNA/RNA into agarose gel and subjecting to current, causing DNA to migrate toward positive pole
nucleic acids with more negative charge migrate more slowly through the matrix
Ion-exchange chromatography
specific region of DNA
minisatellite
phosphorimaging and light detect (chemiluminescence)
tools to evaluate strength of molecular signal (from radioactive or chemical tracers)
denatures proteins and adds negative charge to proteins so they’ll migrate toward the anode
sodium dodecyl sulfate (SDS)
polyacrylamide gel electrophoresis (PAGE)
used to separate/purify polypeptides
Ion-exchange chromatography
nucleic acids with more negative charge migrate more slowly through the matrix
the ability of a strand of DNA/RNA to form a double helix with another strand of DNA/RNA
hybridization
highly sensitive techniques used to detect a gene, nucleic acid, or protein of interest
tracers, labels, probes
southern blotting
invented by Edward Southern; transferred DNA from gel to thin membrane of microcellulose
gel is run and placed in “sandwich” with membrane and exposed to current to transfer DNA. Membrane/filter is then washed, blocked, and exposed to highly specific probe
detection of RNA
blotted from gel to thin membrane of nitrocellulose, then probed with highly specific probe (usually cDNA) for specific RNA of interest
e.g. rat tissue probed for glyceraldehyde-3-phosphate dehydrogenase (G3PHD) [helps make ATP in cells and tissures]
Northern blotting (e.g.)
