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.
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, SE-751 85 Uppsala, Sweden.
Dan-Oscar
DO
Antson
Maritha
M
Mendel-Hartvig
Ulf
U
Landegren
Mats
M
Nilsson
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