Specific primer sequence of cattle Y-chromosome

Abstract

The present invention discloses a specific primer sequence of cattle Y-chromosome and its application in cattle embryo six identification. Cattle ear tissue DNA is first extracted to perform primer specificity identification; and embryo cell sample DNA is then extracted to perform PCR sex identification. The bull sample may be amplified to 255 bp segment when external primer is used in PCR reaction and to 205 bp when internal primer is used, while cow sample has no any amplified product, and this shows the bull specificity of the primer and the applicability of the primer in identifying cattle embryo sex. The present invnetion has the advantages of less pollution, high accuracy and high sensitivity and is suitable for field operation.

Description

technical field [0001] The invention relates to a bovine Y-chromosome-specific primer sequence for identifying the sex of bovine early embryos by means of nested PCR. Background technique [0002] In the field of agricultural biotechnology, sex control, especially the sex control of large livestock such as cattle, has long been one of the major theoretical topics that biological scientists have devoted themselves to research, and has huge economic value. There are two main ways to realize the artificial control of the sex of domestic animals: one is the separation of X sperm and Y sperm; the other is the sex identification of early embryos. However, judging from the current X and Y sperm separation technology, although the use of flow cytometry has successfully separated X and Y sperm, the cost is high and the separation efficiency is low, so it is difficult to promote it in practical applications for a while. With the industrialization of embryo transfer technology, th...

AI Technical Summary

Problems solved by technology

At present, the sequences commonly used to identify the sex of bovine embryos include Y-chromosomal repeat sequences and SRY gene core sequences, etc., but Y-chromosome repeat sequences often affect the accuracy of embryo sex identification because of the possible presence of homologous sequences on other chromosomes The SRY core sequence used by Zeng Yitao and others also has its shortcomings, because the core sequence of the SRY gene has a high homology (up to more than 80%) between mammals, such as the core sequence of the SRY gene of people. The homology is as high as 84%, so that the DNA of male operators and other animals is likely to contaminate the identification process when conducting bovine embryo sex identification, especially when performing PCR operations in the wild, because the environmental conditions are not strict enough, it is easy to produce false positives, Thus affecting the accuracy of embryo sex identification

In addition, the sensitivity of one-time amplification of conventional PCR used by Herr CM and others is limited, because the number of cells that can be cut and sampled from the embryo is very small during embryo sex identification, so sometimes the amount of amplification product is too small to affect the judgment of the identification result

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