Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

Jorrit Schaefer; Goran Jovanovic; Ioly Kotta-Loizou; Martin Buck

doi:10.1016/j.ab.2016.03.017

Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

Jorrit Schaefer, Goran Jovanovic, Ioly Kotta-Loizou, Martin Buck

Research output: Contribution to journal › Article › peer-review

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%.

Original language	English
Pages (from-to)	56-7
Number of pages	2
Journal	Analytical Biochemistry
Volume	503
DOIs	https://doi.org/10.1016/j.ab.2016.03.017
Publication status	Published - 15 Jun 2016
Externally published	Yes

Keywords

Cells, Cultured
Enzyme Assays/instrumentation
Escherichia coli/cytology
beta-Galactosidase/analysis

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10.1016/j.ab.2016.03.017

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title = "Single-step method for β-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 = "Cells, Cultured, Enzyme Assays/instrumentation, Escherichia coli/cytology, beta-Galactosidase/analysis",

author = "Jorrit Schaefer and Goran Jovanovic and Ioly Kotta-Loizou and Martin Buck",

year = "2016",

month = jun,

day = "15",

doi = "10.1016/j.ab.2016.03.017",

language = "English",

volume = "503",

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journal = "Analytical Biochemistry",

issn = "0003-2697",

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T1 - Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

AU - Schaefer, Jorrit

AU - Jovanovic, Goran

AU - Kotta-Loizou, Ioly

AU - Buck, Martin

PY - 2016/6/15

Y1 - 2016/6/15

N2 - 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%.

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%.

KW - Cells, Cultured

KW - Enzyme Assays/instrumentation

KW - Escherichia coli/cytology

KW - beta-Galactosidase/analysis

U2 - 10.1016/j.ab.2016.03.017

DO - 10.1016/j.ab.2016.03.017

M3 - Article

C2 - 27036618

SN - 0003-2697

VL - 503

SP - 56

EP - 57

JO - Analytical Biochemistry

JF - Analytical Biochemistry

ER -

Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

Abstract

Keywords

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