DOI: 10.1101/518118Jan 11, 2019Paper

rEXPAR: an isothermal amplification scheme that is robust to autocatalytic parasites

BioRxiv : the Preprint Server for Biology
Georg UrtelAndré Estévez-Torres

Abstract

In the absence of DNA, a solution containing the four deoxynucleotidetriphosphates (dNTPs), a DNA polymerase and a nicking enzyme generates a self-replicating mixture of DNA species called parasite. Parasites are problematic in template-based isothermal amplification schemes such as EXPAR, as well as in related molecular programming languages, such as the PEN DNA toolbox. Here we show that the nicking enzyme Nb.BssSI allows to change the sequence design of EXPAR templates in a way that prevents the formation of parasites when dATP is removed from the solution. This method allows to make the EXPAR reaction robust to parasite contamination, a common feature in the laboratory, while keeping it compatible with PEN programs, which we demonstrate by engineering a parasite-proof bistable reaction network.

Related Concepts

DNA
DNA-Directed DNA Polymerase
Laboratory
Parasites
Virus Replication
2'-deoxyadenosine triphosphate
DNA Amplification
PSENEN protein, human
Species
Loop-mediated Isothermal Amplification

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