Constrution Of In Situ Hybridiziation For Sperm Specific Expression Gene

Some mRNA sequence of Y-X sperms of deduct hybridization from GaoQingHua proressor of our lab, but differentially expressed genes sequence belongs to Y sperm only in theory. Needing through the experimert method to determine which brlongs to Y sperm drifferentially expressed genes in situ hybridization.Do not any reports about sperm RNA in situ hybridization, because RNA extremely easy to decay. Sperm exist RNA and DNA, DNA removed by Dnae A, and RNA be fixed for hybridization with prob.Sperm DNA and protamine close together, the sperm forms by great destruction use the traditional methods. This experiment used protease K remove protamine from sperm, established a new differentally expressed genes sperm DNA in situ hybridization. Comparison the permeability time of train chromosomes, optimize the chromosomal location technology.The experiment select262bp gene sequence in Y sperm RNA is differentially expressed genes sequence. It prove the Y-X deduct hybridization is successed. The experiment selected323bp gene sequences in chromosome. In37℃, with the30μ g/mL proteinase K concentration processing sperm can remove protamine effectively. The experiment select262bp genes sequence belongs to5chromosomes. Thawing culture medium in38.5℃preheated2h above.it is advantageous for chromosome of cultivation.This experiment is established the RNA differentially expressed genes in situ hybridization. It is established the DNA sperm in situ hybridization. Then this experiment is optimization chromosomal of location technology.