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A Subtractive Hybridization Method to Isolate Tissue-Specific Transcripts: Application to the Selection of New Brain-Specific Products

✍ Scribed by D. Marechal; C. Forceille; D. Breyer; D. Delapierre; A. Dresse

Publisher: Elsevier Science
Year: 1993
Tongue: English
Weight: 373 KB
Volume: 208
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.1993.1055

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✦ Synopsis

A method based on subtractive hybridization of brain complementary DNAs with peripheral messenger RNAs has enabled us to construct an enriched brainspecific 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 doublestranded 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. ic: 1993 Academic Press, Inc.

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