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

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

doi:10.1016/j.xpro.2021.100899

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

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

Facultad de Ciencias de la Salud

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

6 Citas (Scopus)

Resumen

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).

Idioma original	Inglés
Número de artículo	100899
Publicación	STAR Protocols
Volumen	2
N.º	4
DOI	https://doi.org/10.1016/j.xpro.2021.100899
Estado	Publicada - 17 dic. 2021

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abstract = "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 Alc{\'a}ntara et al. (2021a, 2021b).",

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T1 - A low-cost and open-source protocol to produce key enzymes for molecular detection assays

AU - Mendoza-Rojas, Gabriel

AU - Sarabia-Vega, Vanessa

AU - Sanchez-Castro, Ana

AU - Tello, Lesia

AU - Cabrera-Sosa, Luis

AU - Nakamoto, Jose A.

AU - Peñaranda, Katherin

AU - Adaui, Vanessa

AU - Alcántara, Roberto

AU - Milón, Pohl

PY - 2021/12/17

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N2 - 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).

AB - 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).

KW - Biotechnology and bioengineering

KW - CRISPR

KW - Health Sciences

KW - Microbiology

KW - Molecular Biology

KW - Protein Biochemistry

KW - Protein expression and purification

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DO - 10.1016/j.xpro.2021.100899

M3 - Artículo

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SN - 2666-1667

VL - 2

JO - STAR Protocols

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ER -

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

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