Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-05-05
1997-12-02
Chambers, Jasemine C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 912, 4351723, 4353201, 536 243, 536 2431, 536 2433, C12Q 168, C07H 2104, C12P 1934, C12N 1509
Patent
active
056934645
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.
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Meltzer Paul S.
Trent Jeffrey M.
Chambers Jasemine C.
Priebe Scott D.
The Regents of the University of Michigan
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