Hinge-initiated Primer-dependent Amplification of Nucleic Acids (HIP) – A New Versatile Isothermal Amplification Method

  • The growing demand for cost-effective nucleic acid detection assays leads to an increasing number of different isothermal amplification reaction methods. However, all of the most efficient methods suffer from highly complex assay conditions due to the use of complicated primer sets and/or auxiliary enzymes. The present study describes the application of a new linker moiety that can be incorporated between a primer and a secondary target binding site which can act both as a block to polymerase extension as well as a hinge for refolding. This novel “hinge-primer” approach results in an efficient regeneration of the primer binding site and thus improves the strand-displacement and amplification process under isothermal conditions. Our investigations revealed that the reaction with forward and reverse hinge-primer including an abasic site is very efficient. The assay complexity can be reduced by combining the hinge-primer with a corresponding linear primer. Furthermore, the reaction speed can be increased by reducing the length of the amplified target sequence. We tested the sensitivity down to 104 copies and found a linear correlation between reaction time and input copy number. Our approach overcomes the usually cumbersome primer-design and extends the range of isothermal amplification methods using a polymerase with strand-displacement activity.

Download full text files

Export metadata

Additional Services

Search Google Scholar Statistics
Metadaten
Author:Jens Fischbach, Marcus FrohmeORCiDGND, Jörn GlöklerORCiD
URN:urn:nbn:de:kobv:526-opus4-9824
DOI:https://doi.org/10.1038/s41598-017-08067-x
ISSN:2045-2322
Parent Title (English):Scientific Reports
Document Type:Article
Language:English
Year of Publication:2017
Publishing Institution:Technische Hochschule Wildau
Release Date:2017/08/10
Tag:DNA synthesis; Pathogens
Volume:7
Source:Fischbach, J., Frohme, M., & Glökler, J. (2017). Hinge-initiated Primer-dependent Amplification of Nucleic Acids (HIP) – A New Versatile Isothermal Amplification Method Scientific reports. 7, 7683. doi:10.1038/s41598-017-08067-x
Faculties an central facilities:Fachbereich Ingenieur- und Naturwissenschaften
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Licence (German):Creative Commons - CC BY - Namensnennung 4.0 International
Verstanden ✔
Diese Webseite verwendet technisch erforderliche Session-Cookies. Durch die weitere Nutzung der Webseite stimmen Sie diesem zu. Unsere Datenschutzerklärung finden Sie hier.