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BioChip Journal

BioChip Journal

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Inhibition of Non-specific Amplification in Loop-Mediated Isothermal Amplification via Tetramethylammonium Chloride.

Published:1 January 2022 DOI: 10.1007/s13206-022-00070-3 PMID: 35909465
MinJu Jang, Sanghyo Kim

Abstract

Loop-mediated isothermal amplification (LAMP) may be used in molecular and point-of-care diagnostics for pathogen detection. The amplification occurs under isothermal conditions using up to six primers. However, non-specific amplification is frequently observed in LAMP. Non-specific amplification has the potential to be triggered by forward and reverse internal primers. And the relatively low reaction temperature (55-65?°C) induces the secondary structure via primer-primer interactions. Primer redesign and probe design have been recommended to solve this problem. LAMP primers have strict conditions, such as Tm, GC contents, primer dimer, and distance between primers compared to conventional PCR primers. Probe design requires specialized knowledge to have high specificity for a target. In polymerase chain reaction (PCR), some chemicals or proteins are used for improving specificity and efficiency. Therefore, we hypothesized that additives can suppress the non-specific amplification. In this study, tetramethylammonium chloride (TMAC), formamide, dimethyl sulfoxide, Tween 20, and bovine serum albumin have been used as LAMP additives. In our study, TMAC was presented as a promising additive for suppressing non-specific amplification in LAMP.

Supplementary information: The online version contains supplementary material available at 10.1007/s13206-022-00070-3.

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Procduct Name CAS Molecular Formula Supplier Price
Dimethyl sulfoxide 67-68-5 C2H6OS 1924 suppliers $5.00-$120900.00
Polysorbate 20 9005-64-5 C26H50O10 1055 suppliers $9.00-$6080.49
Bovine albumin 9048-46-8 N/A 914 suppliers $15.00-$18680.00
Formamide 75-12-7 CH3NO 851 suppliers $7.62-$5150.00
Tetramethylammonium chloride 75-57-0 C4H12ClN 754 suppliers $10.00-$1674.11
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