Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures

Sheng Cai, Cheulhee Jung, Sanchita Bhadra, Andrew D. Ellington

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 °C were comparable with a regular LAMP reaction at 65 °C.

Original languageEnglish
Pages (from-to)8290-8294
Number of pages5
JournalAnalytical chemistry
Volume90
Issue number14
DOIs
Publication statusPublished - 2018 Jul 17

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Amplification
Temperature
DNA-Binding Proteins
Nucleic Acids
Urea
Testing

ASJC Scopus subject areas

  • Analytical Chemistry

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Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures. / Cai, Sheng; Jung, Cheulhee; Bhadra, Sanchita; Ellington, Andrew D.

In: Analytical chemistry, Vol. 90, No. 14, 17.07.2018, p. 8290-8294.

Research output: Contribution to journalArticle

Cai, Sheng ; Jung, Cheulhee ; Bhadra, Sanchita ; Ellington, Andrew D. / Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures. In: Analytical chemistry. 2018 ; Vol. 90, No. 14. pp. 8290-8294.
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