Isothermal Amplification of Long, Discrete DNA Fragments Facilitated by Single-Stranded Binding Protein.

ABSTRACT: Isothermal amplification methods for detection of DNA and RNA targets have expanded significantly in recent years, promising a new wave of simple and rapid molecular diagnostics. Current isothermal methods result in the generation of short fragments (<150 base pairs) or highly branched long DNA products. Here we report the amplification of discrete target fragments of several kilobases at 37 °C from both double- and single-stranded circular template DNA using specific primer pairs. In contrast to existing methods, this amplification requires only the single-stranded DNA-binding protein gp32 from bacteriophage T4 and a strand-displacing DNA polymerase. In addition to the discrete amplicon products, this method also produces higher molecular weight products consisting of multiple repeated co

SUBMITTER: Zhang Y 

PROVIDER: S-EPMC5561150 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature