Detection of microdeletion syndromes by fluorescence in situ hybridization based on degenerate oligonucleotide primed-PCR

Abstract

Objective To optimize the work of probe preparation used for fluorescence in situ hybridization(FISH) and aplly FISH analysis for the detection of deletions 7q1 1.2,15q11.2,and 22q1 1.2.Methods The DNA of bacterial artificial chromosome (BAC) was amplified by degenerate oligonucleotide primed PCR (DOP-PCR).Indirect labeling of the amplified products was performed by nick translation.Subsequently,the prepared FISH probes were used for detection of chromosome microdeletion.Results By means of DOP-PCR,we got sufficient amounts of FISH probes which provided clear hybridization signals and low backgroud.FISH analysis showed one case with of 15 q 11.2 deletion and another case with a mosaic 7q1 1.2 deletion.Conclusion The optimized method developed by us enables generation of probes to run faster without spending much time on isolating BAC-DNA.The probes could be applied in FISH analysis for the detection of microdeletion syndromes. Key words: Degenerate oligonucleotide primed-PCR (DOP-PCR) ; Fluorescence in situhybridization (FISH) ; Microdeletion syndrome ; Mosaic