Techniques for Gene Analysis Flashcards
Restriction endonucleases
bacterial enzymes capable to cutting DNA at specific palindromic sites
restriction fragments
strands of DNA cut by restriction enzymes
gel electrophoresis
uses DNA’s negative charge to separate based on size
pulsed field gel electrophoresis
used for separating very large DNA fragments
Southern blot
a technique using a labeled probe to determine size of specific DNA fragment or location of endonuclease sites
probe
a known sequence of DNA or RNA used to hybridize to DNA, usually is radioactively labeled
autoradiography
technique used to show which DNA band a probe will hybridize to
Restriction map
the location of various restriction sites on a specific sequence of DNA
Detecting Sickle Cell Anemia
because of known mutation there is a loss of a restriction site, using digestion of DNA and southern blot, disease can be confirmed
Restriction fragment length polymorphisms (RFLPs)
used to determine individual differences in DNA by using restriction enzymes and determining size differences
DNA fingerprinting
takes advantages of DNA polymorphisms to specifically identify differences between DNA samples
Fluorescence in situ hybridization (FISH)
uses fluorescent probes to identify the presence or absence or specific DNA sequences on chromosomes, need to denature DNA without destroying it
Northern blot
similar to southern blot but for RNA, gives info on size and amount of specific RNA molecules
Western blot
Similar to southern but for proteins, uses SDS denaturing gel and radiolabeled antibody
DNA band shifts
used to determine DNA-protein binding, if protein is bound, the DNA-protein complex will be slow on the gel resulting in a high band
