Direct cDNA selection with DNA microdissected from mouse chromosome 16

Isolation of novel clones and construction of a partial transcription map of the C3-C4 region

Jianjun Wei, M. E. Hodes, Yi Wang, Yue Feng, Bernardino Ghetti, Stephen Dlouhy

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A group of cDNA segments was selected by direct hybridization of mouse cerebellar cDNAs against genomic DNA pools generated by microdissection of the mouse chromosome 16 (MMU16) C3-C4 region. After elimination of repetitive sequences and adjustment for redundancy among clones, 34 novel cDNA fragments were isolated. The MMU16 origin of clones was confirmed by genetic linkage mapping. Reverse transcription PCR indicated that ~ 68% of the cDNAs represent transcripts that are expressed in adult mouse cerebellum. Northern blotting showed that some of these are predominantly or solely expressed in brain. This work demonstrates that DNA microdissected from banded MMU16 can be used for direct cDNA selection, thus enabling construction of a new, region-specific partial transcription map. This selected cDNA library should be a useful reagent for further molecular neurobiological studies.

Original languageEnglish
Pages (from-to)678-687
Number of pages10
JournalGenome Research
Volume6
Issue number8
StatePublished - Aug 1996

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Chromosomes, Human, Pair 16
Complementary DNA
Clone Cells
Microdissection
DNA
Genetic Linkage
Nucleic Acid Repetitive Sequences
Chromosome Mapping
Gene Library
Northern Blotting
Cerebellum
Reverse Transcription
Polymerase Chain Reaction
Brain

ASJC Scopus subject areas

  • Genetics

Cite this

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title = "Direct cDNA selection with DNA microdissected from mouse chromosome 16: Isolation of novel clones and construction of a partial transcription map of the C3-C4 region",
abstract = "A group of cDNA segments was selected by direct hybridization of mouse cerebellar cDNAs against genomic DNA pools generated by microdissection of the mouse chromosome 16 (MMU16) C3-C4 region. After elimination of repetitive sequences and adjustment for redundancy among clones, 34 novel cDNA fragments were isolated. The MMU16 origin of clones was confirmed by genetic linkage mapping. Reverse transcription PCR indicated that ~ 68{\%} of the cDNAs represent transcripts that are expressed in adult mouse cerebellum. Northern blotting showed that some of these are predominantly or solely expressed in brain. This work demonstrates that DNA microdissected from banded MMU16 can be used for direct cDNA selection, thus enabling construction of a new, region-specific partial transcription map. This selected cDNA library should be a useful reagent for further molecular neurobiological studies.",
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AU - Hodes, M. E.

AU - Wang, Yi

AU - Feng, Yue

AU - Ghetti, Bernardino

AU - Dlouhy, Stephen

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