Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader
Abstract
Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ss-D-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%.
Keywords:
Microplate reader / LacZ / B-Galactosidase (Bgal) / beta-GalactosidaseSource:
Analytical Biochemistry, 2016, 503, 56-57Publisher:
- Academic Press Inc Elsevier Science, San Diego
Funding / projects:
- Biotechnology and Biological Sciences Research Council (BBSRC) [BB/J00717X/1]
- U.K. Medical Research Council (MRC) project grants [MR/M017672/1]
- BBSRC [BB/J00717X/1] Funding Source: UKRI
- MRC [MR/M017672/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/J00717X/1] Funding Source: researchfish
- Medical Research Council [MR/M017672/1] Funding Source: researchfish
DOI: 10.1016/j.ab.2016.03.017
ISSN: 0003-2697
PubMed: 27036618
WoS: 000375895700008
Scopus: 2-s2.0-84964458891
Collections
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Schaefer, Jorrit AU - Jovanović, Goran AU - Kotta-Loizou, Loly AU - Buck, Martin PY - 2016 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/935 AB - Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ss-D-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. PB - Academic Press Inc Elsevier Science, San Diego T2 - Analytical Biochemistry T1 - Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader EP - 57 SP - 56 VL - 503 DO - 10.1016/j.ab.2016.03.017 ER -
@article{ author = "Schaefer, Jorrit and Jovanović, Goran and Kotta-Loizou, Loly and Buck, Martin", year = "2016", abstract = "Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ss-D-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%.", publisher = "Academic Press Inc Elsevier Science, San Diego", journal = "Analytical Biochemistry", title = "Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader", pages = "57-56", volume = "503", doi = "10.1016/j.ab.2016.03.017" }
Schaefer, J., Jovanović, G., Kotta-Loizou, L.,& Buck, M.. (2016). Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader. in Analytical Biochemistry Academic Press Inc Elsevier Science, San Diego., 503, 56-57. https://doi.org/10.1016/j.ab.2016.03.017
Schaefer J, Jovanović G, Kotta-Loizou L, Buck M. Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader. in Analytical Biochemistry. 2016;503:56-57. doi:10.1016/j.ab.2016.03.017 .
Schaefer, Jorrit, Jovanović, Goran, Kotta-Loizou, Loly, Buck, Martin, "Single-step method for beta-galactosidase assays in Escherichia coli using a 96-well microplate reader" in Analytical Biochemistry, 503 (2016):56-57, https://doi.org/10.1016/j.ab.2016.03.017 . .