Hinweis (Dez. 2019): PubLister wird demnächst abgelöst. Publikationen der Technischen Hochschule Wildau werden zukünftig in HISinOne nachgewiesen. In diesem Zusammenhang kann es momentan zu Überarbeitungen und zu temporären Veränderungen Ihrer persönlichen Publikationslisten kommen. Bei weiteren Fragen wenden Sie sich bitte an friederike.borchert [at] th-wildau [dot] de.

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

Title:
Hinge-initiated Primer-dependent Amplification of Nucleic Acids (HIP) – A New Versatile Isothermal Amplification Method
Abstract:
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.
Year:
2017
Publication type:
Journal article
Journal:
Scientific reports
ISSN:
2045-2322
Volume:
7
Pages:
7683
Language:
English
Document status:
Open access
PubListerURL:
https://publister.bib.th-wildau.de/publister/public/publication/2343
Keywords:
DNA synthesis, Pathogens
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.
@article{2343,
    author           = {Fischbach, Jens and Frohme, Marcus and Glökler, Jörn Felix},
    title            = {Hinge-initiated Primer-dependent Amplification of Nucleic Acids (HIP) – A New Versatile Isothermal Amplification Method},
    journal          = {Scientific reports},
    year             = {2017},
    volume           = {7},
    pages            = {7683},
    publisher        = {Springer Nature},
    doi              = {10.1038/s41598-017-08067-x},
}


Search this title in Google Scholar