Development of DNA libraries specific for Chinese hamster chromosomes 3, 4, 9, 10, X, and Y by DOP-PCR

In a previous study of ours, DNA libraries specific for Chinese hamster chromosomes 1, 2, 3, 5, 6, 7, and 8 were constructed by single laser flow sorting and the linker adapter PCR method. Since a single laser flow sorting of chromosomes is based on chromosome size, it is difficult to separate chromosomes of equal size, such as chromosome 3 and the X chromosome. This problem can be circumvented by bivariate flow sorting. In the present study, we have used bivariate flow sorting of Chinese hamster chromosomes and the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) for generating DNA libraries specifically for chromosomes 3, 4, 9, 10, X, and Y. Fluorescence in situ hybridization (FISH) analyses indicate that the probe libraries are highly specific and can be applied to various cytogenetic studies.