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

详细信息    查看全文

• 作者：Jorrit Schaefer ; Goran Jovanovic ; Ioly Kotta-Loizou ; Martin Buck ; ^{m.buck@imperial.ac.uk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：LacZ ; B-Galactosidase (Bgal) ; β-Galactosidase ; Microplate reader
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 June 2016
• 年：2016
• 卷：503
• 期：Complete
• 页码：56-57
• 全文大小：361 K

文摘

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%.