title: Recombinase-based isothermal amplification of nucleic acids with self-avoiding molecular recognition systems (SAMRS).
abstract: Recombinase polymerase amplification (RPA) is an isothermal method to amplify nucleic acid sequences without the temperature cycling that classical PCR uses. Instead of using heat to denature the DNA duplex, RPA uses recombination enzymes to swap single-stranded primers into the duplex DNA product; these are then extended using a strand-displacing polymerase to complete the cycle. Because RPA runs at low temperatures, it never forces the system to recreate base-pairs following Watson-Crick rules, and therefore it produces undesired products that impede the amplification of the desired product, complicating downstream analysis. Herein, we show that most of these undesired side products can be avoided if the primers contain components of a self-avoiding molecular recognition system (SAMRS). Given the precision that is necessary in the recombination systems for them to function biologically, it is surprising that they accept SAMRS. SAMRS-RPA is expected to be a powerful tool within the range of amplification techniques available to scientists.
authors.affiliation: Foundation for Applied Molecular Evolution, P.O. Box 13174, Gainesville FL 32604 (USA).
authors.forename: Nidhi
authors.initials: N
authors.lastname: Sharma
authors.forename: Shuichi
authors.initials: S
authors.lastname: Hoshika
authors.forename: Daniel
authors.initials: D
authors.lastname: Hutter
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Bradley
authors.forename: Steven A
authors.initials: SA
authors.lastname: Benner
mesh_terms: D009696:Nucleic Acids / Q000096:biosynthesis* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D045522:Recombinases / Q000378:metabolism*; D013696:Temperature
chemical_list: D009696:Nucleic Acids; D045522:Recombinases
keywords: isothermal amplification; nucleotide analogues; oligonucleotides; polymerase chain reaction; recombinase polymerases