Ultra-rapid multicolor PRINS protocol for chromosome detection in human sperm.
We report a new multicolor PRINS procedure for chromosome identification on human sperm. Based on the direct in-situ mixing of the colors of the fluorochromes (FITC, TRITC, Cascade Blue) incorporated in sequential PRINS reactions, this method facilitates rapid distinct labeling of 3 or 4 chromosomes. Each PRINS reaction consists of a unique 4 minute step for annealing and elongation. The method was successfully tested on lymphocytes and spermatozoa. Estimates of disomy were performed for chromosomes 7, 9 and 16 on sperm samples from 2 healthy donors. There was no significant difference between the disomy rates obtained with the conventional two-color PRINS technique and this new three-color procedure. By simplifying the multicolor PRINS protocol, this new protocol should facilitate the use and adaptation of PRINS to various cytogenetic applications.
Pellestor
F
F
CNRS-UPR 1142, Montpellier, France. Franck.Pellestor@igh.cnrs.fr
Malki
S
S
Andréo
B
B
Lefort
G
G
D002885:Chromosomes, Human, Pair 16; D002897:Chromosomes, Human, Pair 7; D002899:Chromosomes, Human, Pair 9; D017931:DNA Primers; D004171:Diploidy; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008540:Meiosis; D008856:Microscopy, Fluorescence; D020401:Primed In Situ Labeling / Q000379:methods*; D013094:Spermatozoa / Q000378:metabolism*
D017931:DNA Primers