Maréchal D, Forceille C, Breyer D, Delapierre D, Dresse A
Laboratoire de Pharmacologie, Université de Liège, Sart Tilman, Belgium.
Anal Biochem. 1993 Feb 1;208(2):330-3. doi: 10.1006/abio.1993.1055.
A method based on subtractive hybridization of brain complementary DNAs with peripheral messenger RNAs has enabled us to construct an enriched brain-specific cDNA library. Single-stranded cDNAs (ssc DNAs) were synthesized from brain polyadenylated mRNAs and subsequently hybridized with peripheral mRNAs immobilized on nitrocellulose membrane. Unhybridized sscDNAs were converted into double-stranded cDNAs and cloned into plasmid pUC13. The screening of the resulting library showed that a high percentage of the cloned cDNAs corresponded to mRNAs specifically transcribed in the brain.
一种基于脑互补DNA与外周信使RNA消减杂交的方法，使我们能够构建一个富集的脑特异性cDNA文库。从脑多聚腺苷酸化mRNA合成单链cDNA（sscDNA），随后与固定在硝酸纤维素膜上的外周mRNA杂交。未杂交的sscDNA转化为双链cDNA并克隆到质粒pUC13中。对所得文库的筛选表明，高比例的克隆cDNA对应于在脑中特异性转录的mRNA。
