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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bsefnbbn

Authors: Tamara Matute, Isaac Núñez, Fernan Federici

Group: Laboratorio de Tecnologias Libres, Reclone.org (The Reagent Collaboration Network)

Summary: This protocol describes how to perform LAMP / RT-LAMP reactions with home-made reagents, which means home-made buffers and positive controls, together with home-brewed BstLF and MMLV enzymes.Home-brewed enzymes information:BstLF polymerase:expression and purification protocol. expression plasmid sequence.M-MLV reverse transcriptase:expression and purification protocol. expression plasmid sequence. Positive control informationSample preparation protocol here.TestingWe test this protocol in LAMP or RT-LAMP reactions with SARS-CoV-2 synthetic dsDNA or ssRNA respectively. As shown in Figure 1, the general performance of reactions with homemade reagents seems appropriate compared with the commercial standard option. No amplification was observed in non-template controls (NTC).[1] Zhang, Y. et al. Enhancing colorimetric loop-mediated isothermal amplification speed and sensitivity with guanidine chloride. Biotechniques69, (2020).

Affiliations: Laboratorio Tecnologías Libres, SynBioUC, Laboratorio Tecnologías Libres

Version: 1

Publication Date: 2021

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