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A subtractive hybridization method to isolate tissue-specific transcripts: application to the selection of new
D Maréchal1, C Forceille, D Breyer
1Laboratoire de Pharmacologie, Université de Liège, Sart Tilman, Belgium.
Analytical Biochemistry
|February 1, 1993
Abstract:
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.