一种简便的高通量细胞内GFP报告基因检测方法

doi:10.3969/j.issn.1004-616x.2015.05.014

Abstract

Abstract:

OBJECTIVE: To establish a simple, fast and high throughput screening method for green fluorescent protein (GFP) assay. METHODS:We applied TECAN multi function measuring instrument infinite M200pro to examine the fluorescence intensity of GFP sample in different dilution buffers, and established the standard curve between fluorescence intensity and GFP relative content. Compared the results obtained from multi function measuring instrument, fluorescence-activated cell sorting (FACS) and Western-blotting. RESULTS:50 mmol/L Tris-HCl (pH=10) was more suitable for examining GFP samples with low content than other dilution buffers. The regression coefficient R2 between fluorescent value and GFP content was above 0.99 by using multi function measuring instrument. The results obtained from multi function measuring instrument correlated significantly with those obtained from FACS and Western-blotting. CONCLUSION:The established method of GFP assay using multi function measuring instrument was convenient, fast and could be used to screen large samples in a short time.

Key words: green fluorescent protein, reporter gene, high throughput screening, multi function measuring instrument

CLC Number:

Q291

YANG Shaozhe, SHI Haijun, CHU Xinran, ZHOU Xiaoling, SUN Pingnan. A simple and high throughput screening method for GFP reporter assay in cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(5): 394-398,403.

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A simple and high throughput screening method for GFP reporter assay in cells

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