Real-time PCR Components

This version of the course is no longer available.
Need multiple seats for your university or lab? Get a quote
The page below is a sample from the LabCE course . Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

Learn more about (online CE course)
Real-time PCR Components

To perform Real-time PCR, the extracted nucleic acid (NA) is combined with individual reagent components necessary to drive the reaction. These components consist of the NA extract along with primers, dNTPs (purine and prymidine bases), and a polymerase enzyme. The mixture is optimized with buffer and additional reagents to stabilize and drive the reaction.
Description of reagents:
  • Primers – Short fragments of oligodeoxynucleotides, 18-25 bps in length, that flank the target sequence.
  • Thermostable Polymerase – Responsible for extension.
  • Enzyme Cofactors – Works with the polymerase, generally magnesium.
  • Free dNTPs – The actual building blocks for DNA.
  • Stabilizer – Includes buffer, glycerol, KOH, etc. stabilizes the reaction.