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

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 (sscDNAs) were synthesized from brain polyadenylated mRNAs and subsequently hybridized with peripheral mRN...

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Bibliographic Details
Published in:Analytical biochemistry 1993-02, Vol.208 (2), p.330-333
Main Authors: Marechal, D., Forceille, C., Breyer, D., Delapierre, D., Dresse, A.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Brain - metabolism
Diverse techniques
DNA - genetics
Evaluation Studies as Topic
Fundamental and applied biological sciences. Psychology
Gene Library
Male
Molecular and cellular biology
Nucleic Acid Hybridization - methods
Organ Specificity
Rats
Rats, Wistar
RNA, Messenger - genetics
Transcription, Genetic
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Summary: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 (sscDNAs) 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.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1993.1055