Recombinase Polymerase Amplification and SIBA

Recombinase polymerase amplification (RPA) and strand-invasion based amplification (SIBA) are isothermal amplification methods enabled through the activity of a recombinase enzyme which help primers invade into double-stranded DNA. T4 UvsX DNA Recombinase is used in combination with its accessory protein, UvsY, and the single-stranded binding protein gp32 to form D-loop recombination structures for initiation of amplification by Bsu or other mesophilic strand-displacing DNA polymerase. RPA uses two oligonucleotides as forward and reverse primers like an “isothermal PCR” while SIBA also includes a longer invasion oligo to help facilitate the strand invasion and amplification. Uniquely, among the isothermal methods RPA can produce amplicons up to 1 kb (primer-to-primer distance). Both RPA and SIBA are typically done at ~37 °C, avoiding the higher temperatures of other methods, though nonspecific amplification can be a common challenge. Probes can be used with either method to detect specific products.  In addition to diagnostic amplification applications, recombinase-based amplification has shown utility for clonal amplification in next generation sequencing workflows.

For information on how we can support your T4 UvsX DNA Recombinase or T4 UvsY Protein needs, contact the OEM & Customized Solutions team.

 

Reaction Temperature Amplicon Size Detection Method(s)
 37°C  <1,000 nt  Fluorescence, Lateral flow

 
Recombinase polymerase amplification workflow

RPA utilizes a recombinase-primer complex, a strand-displacing polymerase, and single-stranded DNA binding proteins to facilitate amplification of discrete DNA products up to 1 kb (primer-to-primer distance).

 


Choose Type:

Legal Information

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.