Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1216, Issue 45, Pages 7670-7676Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2009.09.028
Keywords
CBD-EGFP; CBD-SA; Eukaryotic cells and tissues; Magnetic cellulose microspheres (MCMS); Polyadenylated mRNA isolation
Funding
- National Key Basic Research and Development Program of China [2007CB914803]
- Natural Science Foundation of Tianjin [08JCYBJC04400]
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We have developed micron-sized magnetic cellulose microspheres (MCMS), biospecifically coated with streptavidin (SA) using a novel cellulose binding domain and SA fusion protein, to harvest mRNA from eukaryotic cells and tissues. Biospecific connection between SA and MCMS exhibited significant advantages compared with traditional chemical coupling, including convenient and simple preparation, elimination of toxic compounds, and highly efficient extraction of a pure target molecule. To confirm compatibility, mRNA isolated from cell or tissue samples was used for amplification of housekeeping genes (GAPDH and beta-actin) and specific target genes (two fragments V-H and V-L from the variable fragments of IgG and the gene encoding GLP-1 receptor). Preliminary studies predict that this novel method has potential in future developments in target gene isolation and DNA manipulation. (C) 2009 Elsevier B.V. All rights reserved.
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