Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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

Mendoza-Rojas, Gabriel ; Sarabia-Vega, Vanessa ; Sanchez-Castro, Ana ; Tello, Lesia ; Cabrera-Sosa, Luis ; Nakamoto, Jose A LU orcid ; Peñaranda, Katherin ; Adaui, Vanessa ; Alcántara, Roberto and Milón, Pohl (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:
author
; ; ; ; ; ; ; ; and
publishing date
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}},
}