Chromosomal localization of single copy genes SRY and SOX3 by primed in situ labeling (PRINS).
Primed in situ labeling (PRINS) is a sensitive and specific technique that can be used for the localization of single copy genes and DNA segments that are too small to be detected by conventional FISH. With PRINS, we physically localized the SRY gene to Yp11.31p11.32 and the SOX3 gene to Xq26q27. Locus-specific oligonucleotide primers were annealed in situ and extended on chromosome preparations fixed on microscope slides, in the presence of dATP, dCTP, dGTP, dTTP, biotin-16-dUTP, Tris-HCl, KCl, MgCl2, BSA, and Taq DNA polymerase. Fluorescent signals were detected in metaphase spreads and interphase nuclei. Our method may prove valuable for use with single copy genes in general.
Kadandale
J S
JS
Department of Pediatrics, University of Tennessee, Memphis 38105, USA.
Tunca
Y
Y
Tharapel
A T
AT
D002503:Centromere / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D003854:Deoxyribonucleotides / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D006609:High Mobility Group Proteins / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D009687:Nuclear Proteins; D020161:Physical Chromosome Mapping / Q000379:methods; D020401:Primed In Situ Labeling / Q000379:methods*; D055748:SOXB1 Transcription Factors; D051876:Sex-Determining Region Y Protein; D019914:Taq Polymerase / Q000378:metabolism; D014157:Transcription Factors; D014960:X Chromosome / Q000235:genetics*; D014998:Y Chromosome / Q000235:genetics*
D017931:DNA Primers; D004268:DNA-Binding Proteins; D003854:Deoxyribonucleotides; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; C527785:SOX3 protein, human; D055748:SOXB1 Transcription Factors; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors; D019914:Taq Polymerase