Thermo Fisher Scientific Logo
      Video home
      close menu
      Video home
      close
      • Home
      • Antibodies Videos
      • Bioprocessing Videos
      • Cancer Research Videos
      • Cell Culture Videos
      • Cell & Gene Therapy Videos
      • Clinical Microbiology Videos
      • Digital Science Videos
      • DNA & RNA Extraction & Analysis Videos
      • Electron Microscopy
      • Environmental Videos
      • Food and Beverage Videos
      • Genome Editing Videos
      • Manufacturing & Processing Videos
      • Mass Spectrometry Videos
      • PCR Videos
      • Protein Biology Videos
      • Radiation Detection & Measurement Videos
      • Real-Time PCR Videos
      • Sequencing Videos
      • Spectroscopy, Elemental & Isotope Analysis Videos
      • Stem Cell Research

      Principles of PCR

      1:44

      • Share on Facebook
      • Share on X
      • Share on LinkedIn
      • Share via Email
      Description
      Related Videos

      These reaction components go through multiple cycles of the three main steps of PCR. Denaturation is the first step in the PCR reaction. The sample is heated at a high temperature between 94 and 98 degrees Celcius to separate the double stranded DNA into single strands. The next step is annealing at a temperature between 55 and 72 degrees Celcius, where the primers bind specifically to complementary sequences of both strands of the DNA template. The primers are designed to bracket the region that is targeted for amplification. The optimal annealing temperature for a particular primer pair can be determined experimentally by testing a range of temperatures around 3 to 5 degrees lower than the lowest melting temperature of the two primers. The final step is extension at 68 degrees Celcius or 72 degrees Celcius, depending on the enzyme’s optimal temperature. In this step, the enzyme extends the primer molecules by incorporation of the building blocks, the dNTPs. The polymerase can only start to replicate on the template where a primer has annealed to it. That means that the primer sequence is crucial for amplification of the correct part of the DNA. In some cases, the annealing and extension steps can be combined. This is known as a 2-step cycling protocol.

      View More
      View Less
      Learn more about PCR

      Share

      • Share on Facebook
      • Share on X
      • Share on LinkedIn
      • Share via Email
      •  
      •  
      •  
      •  
      •  
      Brands
      • Thermo Scientific
      • Applied Biosystems
      • Invitrogen
      • Fisher Scientific
      • Unity Lab Services
      Shopping Tool
      • Product Selection Guides - US
      • Quick Order - US
      • Redeem a Quote
        * Please enter a valid quote
      • New Products
      • Promotions
      • Mobile & Desktop Apps
      • Shared Lists
      eSolutions
      • eProcurement
      • Supply Center
      • Instrument Management
      Support
      • Order Support
      • Training
      • Webinars
      • Blog
      • Social Media
      • Contact Us
      • Report a Site Issue
      Thermo Fisher Scientific
      • About Us
      • Careers
      • Investors
      • News
      • Responsibility
      • Trademarks
      United States
      • Terms & Conditions
      • Privacy Information Center
      • Price & Freight Policy

       

      Note: You clicked on an external link, which has been disabled in order to keep your shopping session open.

      Ok

      Share this video

      Embed

      Size: x pixels