Method and kit for identifying milk cattle embryo gender

Abstract

The invention discloses a method and a reagent kit for performing sex appraisal of milk cow embryos by utilizing a real-time fluorescent PCR. A pair of milk cow specific primers is adopted to amplify a 182bp area on a sex determining gene (sry), and a specific TaqMan hydrolysis probe method is used for real-time detection. Components of the reagent kit comprise a nucleic acid concentrate, a nucleic acid extraction liquid, a PCR reaction buffer liquid, Taq enzyme, negative control, and positive control. The use of the reagent kit comprises two steps of sample treatment and amplification detection, the method judges the sex of a sample individual by detecting the existence of a sry gene segment in a template, and the reagent kit has simple, convenient and quick (within 1 hour) operation, can avoid the false positive produced by the nucleic acid pollution of self-amplification products, persons and other animal sources, has the accuracy rate high up to 100 percent, and can be widely applied to the sex appraisal of fetal bovines of pregnant milk cows, the sex appraisal of embryo transplantation of milk cows, and the detection of sry genes of milk cows for scientific research.

Description

technical field [0001] The invention belongs to a method and a kit for detecting the sex of cow embryos by real-time fluorescent PCR. Background technique [0002] Since the advent of the polymerase chain reaction (Polymerase Chain Reaction, referred to as PCR) jointly created by Cetus Corporation of the United States and the University of California in 1985, this process has been performed in the presence of template DNA, primers, deoxyribonucleoside triphosphates, and DNA polymerase. The technology of in vitro enzymatic amplification of DNA immediately aroused widespread interest and attention, and has been widely used and developed around the world, and quickly entered molecular biology, forensic science, archaeology, and nucleic acid detection of genetic diseases and infectious pathogens and other fields. At present, the traditional "PCR+electrophoresis" or "PCR+probe hybridization" detection method has been widely used in the field of nucleic acid detection. However, ...

AI Technical Summary

Problems solved by technology

[0006] However, the main problem of the above-mentioned gender identification technology and kits is that the laboratory conditions and the technical level of operators are relatively high, otherwise false positive results will appear and the detection accuracy will be reduced; secondly, the kits are cumbersome to operate and the time required Longer; finally, the kit can only identify embryonic cells to be transplanted and cannot be used for fetal sex identification in pregnant dairy cows

Due to the above-mentioned limitations, it is not suitable for popularization and use as a mature technology

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