A low-cost and open-source protocol to produce key enzymes for molecular detection assays

Summary: Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcrip...

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Autores principales:	Gabriel Mendoza-Rojas, Vanessa Sarabia-Vega, Ana Sanchez-Castro, Lesia Tello, Luis Cabrera-Sosa, Jose A. Nakamoto, Katherin Peñaranda, Vanessa Adaui, Roberto Alcántara, Pohl Milón
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Health Sciences Microbiology Molecular Biology CRISPR Protein Biochemistry Protein expression and purification Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/ea9665530c934376806cf99d74794974
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spelling	oai:doaj.org-article:ea9665530c934376806cf99d747949742021-11-04T04:39:47ZA low-cost and open-source protocol to produce key enzymes for molecular detection assays2666-166710.1016/j.xpro.2021.100899https://doaj.org/article/ea9665530c934376806cf99d747949742021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006055https://doaj.org/toc/2666-1667Summary: Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcriptase, and TEV protease to make it compatible with minimal laboratory equipment, broadly available in low- and middle-income countries. The enzymes produced with this protocol have been successfully used for molecular detection applications.For complete details on the use and execution of this protocol, please refer to Alcántara et al. (2021a, 2021b).Gabriel Mendoza-RojasVanessa Sarabia-VegaAna Sanchez-CastroLesia TelloLuis Cabrera-SosaJose A. NakamotoKatherin PeñarandaVanessa AdauiRoberto AlcántaraPohl MilónElsevierarticleHealth SciencesMicrobiologyMolecular BiologyCRISPRProtein BiochemistryProtein expression and purificationScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100899- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Health Sciences Microbiology Molecular Biology CRISPR Protein Biochemistry Protein expression and purification Science (General) Q1-390
spellingShingle	Health Sciences Microbiology Molecular Biology CRISPR Protein Biochemistry Protein expression and purification Science (General) Q1-390 Gabriel Mendoza-Rojas Vanessa Sarabia-Vega Ana Sanchez-Castro Lesia Tello Luis Cabrera-Sosa Jose A. Nakamoto Katherin Peñaranda Vanessa Adaui Roberto Alcántara Pohl Milón A low-cost and open-source protocol to produce key enzymes for molecular detection assays
description	Summary: Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcriptase, and TEV protease to make it compatible with minimal laboratory equipment, broadly available in low- and middle-income countries. The enzymes produced with this protocol have been successfully used for molecular detection applications.For complete details on the use and execution of this protocol, please refer to Alcántara et al. (2021a, 2021b).
format	article
author	Gabriel Mendoza-Rojas Vanessa Sarabia-Vega Ana Sanchez-Castro Lesia Tello Luis Cabrera-Sosa Jose A. Nakamoto Katherin Peñaranda Vanessa Adaui Roberto Alcántara Pohl Milón
author_facet	Gabriel Mendoza-Rojas Vanessa Sarabia-Vega Ana Sanchez-Castro Lesia Tello Luis Cabrera-Sosa Jose A. Nakamoto Katherin Peñaranda Vanessa Adaui Roberto Alcántara Pohl Milón
author_sort	Gabriel Mendoza-Rojas
title	A low-cost and open-source protocol to produce key enzymes for molecular detection assays
title_short	A low-cost and open-source protocol to produce key enzymes for molecular detection assays
title_full	A low-cost and open-source protocol to produce key enzymes for molecular detection assays
title_fullStr	A low-cost and open-source protocol to produce key enzymes for molecular detection assays
title_full_unstemmed	A low-cost and open-source protocol to produce key enzymes for molecular detection assays
title_sort	low-cost and open-source protocol to produce key enzymes for molecular detection assays
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/ea9665530c934376806cf99d74794974
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A low-cost and open-source protocol to produce key enzymes for molecular detection assays

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