title: Primed in situ labeling for detecting single-copy genes.
abstract: In  order to analyze male sterility caused by deletion of SRY and DAZ, we examined  the accuracy and cost-effectiveness of a modified primed in situ labeling  (PRINS) technique for detection of single-copy genes. Peripheral blood samples  were collected from 50 healthy men; medium-term cultured lymphocytes from these  samples were suspended in fixative solution and then spread on clean slides. We  used four primers homologous to unique regions of the SRY and DAZ regions of  the human Y-chromosome and incorporated reagents to increase polymerase  specificity and to enhance the hybridization signal. PRINS of SRY and DAZ gave  bands at Yp11.3 and Yq11.2, respectively, in all 50 metaphase spreads. The  PRINS SRY signals were as distinct as those obtained using traditional  fluorescence in situ hybridization (FISH). This new method is ideal for  rapid localization of single-copy genes or small DNA segments, making PRINS a  cost-effective alternative to FISH. Further enhancement of PRINS to increase  its speed of implementation may lead to its wide use in the field of medical  genetics.
authors.affiliation: Key Laboratory of Bio-Resources and Eco-Environment, Institute of Medical Genetics, College of Life Science, Sichuan University, Ministry of Education, Chengdu, China.
authors.forename: J-L
authors.initials: JL
authors.lastname: Gao
authors.forename: Y
authors.initials: Y
authors.lastname: Nie
authors.forename: X-P
authors.initials: XP
authors.lastname: Ding
mesh_terms: D002869:Chromosome Aberrations; D041322:Chromosomes, Human, Y / Q000235:genetics; D017931:DNA Primers; D000075364:Deleted in Azoospermia 1 Protein; D018628:Gene Dosage; D024282:Genes, sry; D006059:Gonadal Dysgenesis / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007248:Infertility, Male / Q000235:genetics*; D008214:Lymphocytes; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D020401:Primed In Situ Labeling / Q000379:methods*; D016601:RNA-Binding Proteins / Q000235:genetics*; D051876:Sex-Determining Region Y Protein / Q000235:genetics*; D013094:Spermatozoa / Q000166:cytology / Q000254:growth & development
chemical_list: C095668:DAZ1 protein, human; D017931:DNA Primers; D000075364:Deleted in Azoospermia 1 Protein; D016601:RNA-Binding Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein