Polymerase Chain Reaction

Overview

Amplifies specific DNA fragments into billions of copies that can be detected in an end-point analysis.

PCR (polymerase chain reaction) is a powerful technique to exponentially amplify DNA regions of interest. It utilizes the ability of naturally occurring DNA polymerases to extend short single-stranded primers that hybridize to the target template DNA. The amplified PCR product can be detected and analyzed using gel electrophoresis and image analysis. PCR is considered the cornerstone of modern molecular biology, with applications ranging from cloning and sequencing to diagnostics and forensic analysis. PCR is a staple technique of every molecular biology lab with sophisticated instruments and reagents available to carry out the reactions efficiently. Most commercially available PCR reagents contain wild-type enzymes, isolated from nature without modification, and depend on factors like buffer formulation or enzyme concentration to provide differences in performance. Roche offers PCR reagents containing DNA polymerase and other enzymes selected through our directed evolution technology to deliver high-quality results for specific PCR applications.

 

What are the advantages of using PCR reagents selected through directed evolution?

  • Contain novel enzymes that are optimized specifically for qPCR application
  • Confer significant improvements to reaction efficiency, sensitivity, speed and signal-to-noise ratio compared to wild-type reagents

 

How does directed evolution work?

Directed evolution exploits the principle of natural selection. Mutagens are used to introduce random variations in the gene encoding DNA polymerase, which results in a library consisting of millions of genes coding for unique variants of the enzyme. Next, a selection pressure is applied to identify the genes that survive the pressure, and those genes are then selected. The process is repeated until the enzymes exhibiting the highest improvements are obtained.

 

Available Products by Application

Roche offers PCR reagents optimally suited for a variety of applications.

Application

Product

Features and Benefits

Cloning

KAPA HiFi PCR Kits

KAPA Long Range PCR Kits

  • High-fidelity amplification of inserts up to 10 kb with an error rate 100X lower than Taq DNA polymerase. Compatible with blunt-end cloning
  • Amplification of inserts up to 15 kb with high sensitivity and an error rate 4X lower than Taq DNA polymerase. Compatible with TA cloning

 

Colony PCR

KAPA2G Robust PCR Kits

KAPA HiFi HotStart PCR Kits

  • Improved tolerance to PCR inhibitors results in higher success rates and streamlined workflows with common E. coli and S. cerevisae strains*
  • Recommended for colony PCR when high fidelity and/or longer targets up to 10 kb are required

 

Crude Sample PCR

KAPA2G Robust PCR Kits

KAPA3G Plant PCR Kits

KAPA Mouse Genotyping Kits

  • Improved resistance to many inhibitors commonly found in crude samples—such as denatured protein, salts and metabolites—results in higher PCR success rates and more streamlined workflows*
  • Improved resistance to inhibitors co-purified in plant DNA extracts, and amplification directly from plant tissues enable higher-throughput workflows and eliminate the need for expensive and time-consuming purification*
  • Increased throughput and decreased turnaround time*
  • Better performance compared to other available crude extraction methods *

 

Direct Plant PCR

KAPA3G Plant PCR Kits
  • Improved resistance to inhibitors co-purified in plant DNA extracts, and amplification directly from plant tissues enable higher-throughput workflows and eliminate the need for expensive and time-consuming purification*

 

Fast PCR

KAPA2G Fast PCR Kits
  • Higher specific activity and processivity significantly reduces PCR cycling times by 50% without a loss in performance*

 

GC-rich PCR

KAPA2G Robust PCR Kits

KAPA HiFi PCR Kits

 

  • Recommended for the amplification of GC-rich targets

 

Genotyping

KAPA2G Robust PCR Kits

KAPA2G Fast PCR Kits

KAPA Taq PCR Kits

KAPA Mouse Genotyping Kits

  • Robust performance, shorter cycling times, streamlined workflows and higher success rates across a wide variety of template types and amplicons*
  • Reduced PCR cycling times and improved success rates streamline workflows for routine genotyping*
  • High quality and optimal formulation for routine genotyping applications
  • Increased throughput and decreased turnaround time*
  • Better performance compared to other available crude extraction methods*

 

 

 

High-fidelity PCR

KAPA HiFi PCR Kits

KAPA LongRange PCR Kits

  • Error rate 100X lower than Taq DNA polymerase. Higher processivity results in significant improvements to fidelity, sensitivity, target length, and the ability to amplify difficult templates*
  • Recommended for the amplification of longer targets up 15 kb with improved sensitivity and an error rate 4X lower than Taq DNA polymerase

 

Long-range PCR

KAPA LongRange PCR Kits

KAPA HiFi PCR Kits

  • Recommended for the amplification of longer targets up 15 kb with improved sensitivity and an error rate 4X lower than Taq DNA polymerase*
  • High-fidelity amplification of inserts up to 10 kb with an error rate 100X lower than Taq DNA polymerase*

 

Multiplex PCR

KAPA2G Fast Multiplex Kits

  • Increased specific activity and processivity, formulated for highly sensitive and uniform multiplex amplification*

 

Single-protocol PCR

KAPA2G Robust PCR Kits

KAPA2G Fast PCR Kits

  • Robust performance across a wide range of GC- and AT-rich templates streamlines workflows while reducing turnaround times. Recommended for end-point applications using gel electrophoresis
  • Broad coverage of both AT- and GC-rich targets consolidates protocols and reduces cycling times. Recommended for higher sensitivity detection such as capillary electrophoresis and Sanger sequencing workflows

 

Site-directed Mutagenesis

KAPA HiFi PCR Kits
  • Error rate 100X lower than Taq DNA polymerase.* Higher processivity results in significant improvements to fidelity and target length.* Compatible with site-directed mutagenesis protocols

 

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*Data on file.