A low-cost and open-source protocol to produce key enzymes for molecular detection assays
(2021) In STAR Protocols 2(4). p.1-28- 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 Alcántara et al. (2021a, 2021b).
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/71b9a8b6-7400-417a-a8e2-a72bd52ab6f2
- author
- Mendoza-Rojas, Gabriel ; Sarabia-Vega, Vanessa ; Sanchez-Castro, Ana ; Tello, Lesia ; Cabrera-Sosa, Luis ; Nakamoto, Jose A LU ; Peñaranda, Katherin ; Adaui, Vanessa ; Alcántara, Roberto and Milón, Pohl
- publishing date
- 2021
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Chromatography, Affinity, Enzyme Assays, Enzymes/genetics, Escherichia coli/genetics, Molecular Typing, Recombinant Proteins/genetics, Transformation, Bacterial
- in
- STAR Protocols
- volume
- 2
- issue
- 4
- article number
- 100899
- pages
- 1 - 28
- publisher
- Cell Press
- external identifiers
-
- scopus:85118479458
- pmid:34766029
- ISSN
- 2666-1667
- DOI
- 10.1016/j.xpro.2021.100899
- language
- English
- LU publication?
- no
- additional info
- © 2021 The Author(s).
- id
- 71b9a8b6-7400-417a-a8e2-a72bd52ab6f2
- date added to LUP
- 2022-10-13 10:15:59
- date last changed
- 2024-04-04 12:41:15
@article{71b9a8b6-7400-417a-a8e2-a72bd52ab6f2, abstract = {{<p>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).</p>}}, author = {{Mendoza-Rojas, Gabriel and Sarabia-Vega, Vanessa and Sanchez-Castro, Ana and Tello, Lesia and Cabrera-Sosa, Luis and Nakamoto, Jose A and Peñaranda, Katherin and Adaui, Vanessa and Alcántara, Roberto and Milón, Pohl}}, issn = {{2666-1667}}, keywords = {{Chromatography, Affinity; Enzyme Assays; Enzymes/genetics; Escherichia coli/genetics; Molecular Typing; Recombinant Proteins/genetics; Transformation, Bacterial}}, language = {{eng}}, number = {{4}}, pages = {{1--28}}, publisher = {{Cell Press}}, series = {{STAR Protocols}}, title = {{A low-cost and open-source protocol to produce key enzymes for molecular detection assays}}, url = {{http://dx.doi.org/10.1016/j.xpro.2021.100899}}, doi = {{10.1016/j.xpro.2021.100899}}, volume = {{2}}, year = {{2021}}, }