Construction and characterization of a region-specific microdissection library from human chromosome 2q35-q37
| Autor: | Teresa Yang-Feng, Suhong Tong, Jingwei Yu, Fa-Ten Kao |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Genetics
Yeast artificial chromosome Genomic Library medicine.diagnostic_test cDNA library Chromosome Biology Polymerase Chain Reaction Molecular biology Cell Line Blotting Southern Chromosomes Human Pair 2 Chromosome regions medicine Humans Genomic library Cloning Molecular In Situ Hybridization Fluorescence Microdissection Repetitive Sequences Nucleic Acid Southern blot Fluorescence in situ hybridization |
| Zdroj: | Genomics. 14:769-774 |
| ISSN: | 0888-7543 |
| Popis: | A region-specific genomic library for human chromosome 2q35-q37 has been constructed using the microdissection and polymerase chain reaction-mediated linker-adaptor microcloning method. Twenty fragments from the chromosome region 2q35-q37 were dissected and a library consisting of 20,000 recombinant microclones was obtained. The insert size ranged between 50 and 800 bp, with a mean of approximately 270 bp. About 50–60% of the microclones contained unique sequences. The microdissection library has been demonstrated to derive from the dissected region 2q35-q37 by chromosome painting using the fluorescence in situ hybridization (FISH) technique. Southern blot analysis of the unique sequence microclones from the library showed that 54% (26/48) of the clones are of human origin and chromosome 2 specific. Four of these microclones have been further mapped to the 2q37 region by using a cell hybrid containing only 2q37. The unique sequence microclones have also been characterized for their insert size and the hybridizing genomic fragments cleaved with Hin dIII. As shown previously, these microclones will be useful in isolating corresponding yeast artificial chromosome (YAC) clones with large inserts for high-resolution physical mapping and also in screening cDNA libraries to isolate expressed gene sequences as candidate genes to facilitate search for the crucial genes underlying genetic diseases and specific forms of cancer assigned to the region. |
| Databáze: | OpenAIRE |
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