PRINS for the detection of unique sequences.
Primed in situ labeling (PRINS) can be used to localize single-copy genes and unique sequences. Using a modified PRINS method that incorporates multiple primers for the same sequence, single-step annealing and extension, anti-Taq DNA polymerase antibody, and stringent washing, we localized the human SRY gene to Yp11.31-p11.32 in chromosome preparations in situ. Locus-specific oligonucleotide probes (i.e., PRINS primers) were annealed to chromosomal DNA fixed on glass slides and extended in the presence of the four trinucleotide precursors, biotin-16-2'-deoxyuridine 5'-triphosphate, Tris-HCl, KCl, MgCl2, bovine serum albumin, and Taq DNA polymerase. After reaction with avidin-conjugated fluorophores, the resulting signals could be visualized by fluorescence microscopy in metaphase spreads and in interphase nuclei. This method could prove useful for unique sequences in general.
Wachtel
Stephen S
SS
Department of Obstetrics and Gynecology, University of Tennessee, Memphis, TN, USA.
Tharapel
Avirachan T
AT
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D008856:Microscopy, Fluorescence; D020401:Primed In Situ Labeling / Q000379:methods*
D004247:DNA