Method for generating chromosome region-specific probes
US5693464A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 5, 1994 |
| Grant date | Dec 2, 1997 |
| Priority date | — |
| Expiry date | May 5, 2014 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/156
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides rapid, reproducible procedures for generating chromosome region-specific probes (CRSPs) for diagnostic and research applications. Region-specific probes are provided by direct in vitro enzymatic amplification (PCR) of microdissected chromosomal or hybridized DNA from the chromosomal region of interest, followed by labelling for in situ hybridization to metaphase chromosomes and interphase nuclei. CRSP specificity can be further enhanced using a linker-based strategy, wherein linkered DNA (LDNA) sequences prepared from DNA libraries are hybridized to chromosomal DNA in situ, microdissected from the chromosomal region of interest and then directly amplified using the linker as primer. These procedures make it possible to generate a vast number of chromosome region-specific probes without microchemical manipulation after dissection and provide means for identifying cryptic chromosomal alterations previously not amenable to routine cytogenetic analysis. Probes generated by the methods of the present invention can also be used for screening any DNA library of interest.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.