Molecular Diagnosis Flashcards
Why use DNA RNA and proteins to help identify pathogenic agents?
Specificity
Sensitivity
Reduction in dependency on culture
Saftey
__________ is a technique used in order to separate macromolecules based on ______. The technique applies a negative charge so _________ move towards a positive charge.
Structure:
_________>__________>_________(DNA)
_________>__________>_________>___________(Proteins)
Electrophoresis
size
proteins
supercoiled>linear>nicked(DNA)
monomer>dimer>trimer>polymer(proteins)
restriction enzymes
cut DNA at or near specific recognition nucleotide sequences
sequence specific
Hybridization
-examples?
denatured single-stranded DNA probe binds to a complementary single-stranded sequence
Dot, in situ, Southern, Northern, microarray
a hybridization probe can be…
nucleic acid fragment
labeled
radioisotope
enzyme
Target amplification
-examples?
enzyme-mediated process to synthesize copies of targeted nucleic acid
PCR and isothermal amplification
Target amplification has a __________ (pro) but can _________________ (con).
high sensitivity
flase positive
The ____________________ is a technology in molecular biology used to amplify a single copy or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
polymerase chain reaction (PCR)
PCR Primer
single-stranded DNA fragments, complementary to sequences flanking the region to be amplified
A real-time polymerase chain reaction or quantitative polymerase chain reaction is used to ________ and simultaneously __________ or quantify a targeted DNA molecule. Uses a probe or dye to generate a _________ signal from the product.
amplify
detect
fluorescent
What has a higher sensitivity than PCR and gives visible results?
LAMP
