PCR (Polymerase Chain Reaction) detection liquid for loss of loci SY84, SY134 and SY255 of human Y chromosome

Abstract

The invention discloses PCR (Polymerase Chain Reaction) detection liquid for the loss of loci SY84, SY134 and SY255 of a human Y chromosome. The PCR detection liquid comprises magnesium ions, dNTP (deoxy-ribonucleoside triphosphate), Taq enzyme, potassium chloride, tris(hydroxymethyl)aminomethane-hydrochloric acid, a SY84 primer, a SY134 primer and a SY255 primer; and in the PCR detection liquid, the concentration of the magnesium ions is 2.632 mM, the concentration of the dNTP is 0.263 mM, the concentration of the Taq enzyme is 105 u / ml, the concentration of the potassium chloride is 50.63 mM, the concentration of the trisaminomethane-hydrochloric acid with the pH of 8.3 is 10.53 mM, the concentration of the SY84 primer is 0.263 [mu]M, the concentration of the SY134 primer is 0.421 [mu]M and the concentration of the SY255 primer is 0.263 [mu]M. The PCR detection liquid can be stored for a long term at low temperature and can withstand repeated freezing and thawing, only a DNA (Deoxyribonucleic Acid) template is required to be added during detection, the operation is simple, convenient and fast, and the amplification effect is specific and sensitive.

Description

technical field [0001] The invention relates to a PCR detection technology for human Y chromosome deletion, in particular to a PCR detection solution for deletion of human Y chromosome SY84, SY134 and SY255 sites. Background technique [0002] About 10% to 15% of couples of childbearing age are infertile, of which 50% are caused by men. In addition to clear causes such as vas deferens obstruction and infection, changes in genetic traits are also an important reason, mainly due to male spermatogenesis disorders. Manifested as severe oligospermia (less than 2×10 6 / mL~5×10 6 / mL) or no sperm. About 20% of clinically infertile men are caused by hereditary, non-obstructive azoospermia or oligospermia, and most of these patients have normal karyotype tests; but Y chromosome microdeletions lead to hereditary infertility. At present, PCR of STSs is generally used to obtain Y chromosome microdeletion information, but the deletion rate reported in various literatures varies from 1...

AI Technical Summary

Problems solved by technology

When detecting these STS sites by PCR, water, magnesium ions, buffer solutions (KCl and Tris-HCl, etc.), dNTPs, primers, enzymes, etc. need to be gradually added to the reaction system in steps during the operation. The operation process is cumbersome, and each group added Due to the different proportions of the components, the expansion efficiency of nucleic acid is different, and there are problems such as a sharp drop in amplification efficiency when repeated freezing and thawing.