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LAMP/RT-LAMP Reaction protocol v1

Tamara Matúte, Isaac Núñez, Fernán Federici

License: CC BY

Abstract: 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 information Sample preparation protocol here. Testing We 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).

Published on 14 Feb. 2021