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

48 Citations (Scopus)

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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