PCR-generated padlock probes distinguish homologous chromosomes through quantitative fluorescence analysis.
Conventional cytogenetic techniques can distinguish homologous chromosomes in a qualitative manner based upon obvious morphological features or using in situ hybridization methods that yield qualitative data. We have developed a method for quantitative genotyping of single-nucleotide variants in situ using circularizable DNA probes, so-called padlock probes, targeting two different alpha satellite repeat variants present in human chromosome 7 centromeres, and a single-nucleotide variation in alpha satellite repeats on human chromosome 15 centromeres. By using these PCR-generated padlock probes, we could quantitatively distinguish homologous chromosomes and follow the transmission of the chromosomes by in situ analysis during three consecutive generations.
Antson
Dan-Oscar
DO
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, SE-751 85 Uppsala, Sweden.
Mendel-Hartvig
Maritha
M
Landegren
Ulf
U
Nilsson
Mats
M
D002503:Centromere / Q000235:genetics; D002884:Chromosomes, Human, Pair 15; D002897:Chromosomes, Human, Pair 7; D020732:Cytogenetic Analysis / Q000379:methods*; D015342:DNA Probes; D004276:DNA, Satellite; D005260:Female; D005819:Genetic Markers; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D010375:Pedigree; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide
D015342:DNA Probes; D004276:DNA, Satellite; D005819:Genetic Markers