PCR (polymerase chain reaction) detection solution for SY86, SY127 and SY254 site deletions in human Y chromosome

Abstract

The invention discloses a PCR (polymerase chain reaction) detection solution for SY121, SY145 and SY242 site deletions in a human Y chromosome, which comprises magnesium ions, dNTP (deoxyribonucleotide triphosphate), a Taq enzyme, potassium chloride, trihydroxymethyl aminomethane-hydrochloric acid, an SY86 primer, an SY127 primer and an SY254 primer, wherein the concentration of the magnesium ions is 2.105 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 trihydroxymethyl aminomethane-hydrochloric acid with the pH value of 8.3 is 10.53 mM, the concentration of the SY86 primer is 0.263 mu M, the concentration of the SY127 primer is 0.263 mu M, and the concentration of the SY254 primer is 0.158 mu M. The PCR detection solution can be stored at low temperature for a long time, and can resist repeated freeze thawing; when in detection, only a DNA (deoxyribonucleic acid) template needs to be added; and thus, the invention is simple and quick to operate, and has higher specificity and sensitivity.

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 SY86, SY127 and SY254 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 ...

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.