Steps in PCRcontinued
Molecular Diagnostics
(MDx) Review 556
STEP TEMP/TIME* EXPLANATION
- Annealing (hybridization) Primers attach to both template strands by binding with complementary
bases on 3’ side of region to be amplified.
50 °–70°C/
20–90 sec.3 ¢3 ¢DNA
5 ¢5 ¢Primer PrimerRegion of interest(From Buckingham L. Molecular Diagnostics: Fundamentals, Methods, and
Clinical Applications, 2nd ed. Philadelphia: FA Davis; 2012:134.)continued...