Amplification Terminologycontinued
Molecular Diagnostics
(MDx) Review 552
TERM EXPLANATION
Melting temperature (Tm)
Anneal
Polymerase
DNA polymerase
Taq polymerase
Oliogonucleotides
Primers
Deoxynucleotide triphosphates (dNTPs)
Temperature required to denature 50% of dsDNA. Depends in part on GC content. G-C pair
harder to break than A-T pair because 3 H-bonds instead of 2.
Binding to complementary strand of nucleic acid.
Enzyme that assembles nucleotides to produce new strand of nucleic acid.
Enzyme that synthesizes DNA using existing strand of DNA as template. Can only add to
existing piece of DNA so needs primer to start.
Heat-stable DNA polymerase used to catalyze synthesis of DNA in PCR. Originally isolated
from bacterium Thermus aquaticusin hot springs of Yellowstone National Park. Can with-
stand high temp required for denaturation.
Short fragments of ssDNA that are synthesized for specific purpose, e.g., oligonucleotide
primers.
Short segments of DNA designed to anneal to DNA target in PCR. Provide free 3’-OH group to
which DNA polymerase can add dNTP.
Building blocks from which DNA polymerase synthesizes new strands of DNA. Deoxyadenosine
triphosphate (dATP), deoxyguanosine triphosphate (dGTP), deoxythymidine triphosphate
(dTTP), deoxycytidine triphosphate (dCTP).
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