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An improved method with high sensitivity and low background in detecting low β-galactosidase expression in mouse embryos.

ABSTRACT: LacZ is widely used as a reporter in studies of gene expression patterns. β-galactosidase, the product of LacZ gene, is usually detected by X-gal/FeCN staining. In X-gal/FeCN staining, β-galactosidase catalyzes X-gal to produce blue precipitates, which indicate the expression patterns of the gene of interest. A newer LacZ detection method using S-gal/TNBT is more sensitive but plagued by high background. Here, we describe an improved procedure that combines advantageous steps from the two methods. By comparing with X-gal/FeCN and S-gal/TNBT methods in detecting the expression patterns of miR-322/503 and miR-451 at a series of developmental stages, the improved method showed higher sensitivity and lower background. Thus, the improved method could be an alternative way of β-galactosidase staining in low gene expression situations.

SUBMITTER: Shen X

PROVIDER: S-EPMC5419561 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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An improved method with high sensitivity and low background in detecting low β-galactosidase expression in mouse embryos.

Shen Xiaopeng X Bao Wenjing W Yu Wei W Liang Rui R Nguyen Bao B Liu Yu Y

PloS one 20170505 5

LacZ is widely used as a reporter in studies of gene expression patterns. β-galactosidase, the product of LacZ gene, is usually detected by X-gal/FeCN staining. In X-gal/FeCN staining, β-galactosidase catalyzes X-gal to produce blue precipitates, which indicate the expression patterns of the gene of interest. A newer LacZ detection method using S-gal/TNBT is more sensitive but plagued by high background. Here, we describe an improved procedure that combines advantageous steps from the two method ...[more]

PMID: 28475610

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