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Method for detecting sheep PRAMEY gene copy number variation and application thereof

A technology of gene copy number and copy number, which is applied in the field of molecular genetics detection, can solve the problems such as the copy number and variation of sheep PRAMEY gene that have not yet been found, and achieve the effects of high sensitivity, easy operation and low cost.

Pending Publication Date: 2020-07-03
甘肃润牧生物工程有限责任公司
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Problems solved by technology

But up to now, there is no report on the method of detecting the copy number variation of PRAMEY gene in sheep

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  • Method for detecting sheep PRAMEY gene copy number variation and application thereof
  • Method for detecting sheep PRAMEY gene copy number variation and application thereof
  • Method for detecting sheep PRAMEY gene copy number variation and application thereof

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Embodiment Construction

[0031] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. The examples are only used to explain the present invention, not to limit the protection scope of the present invention.

[0032] The invention utilizes the real-time quantitative PCR technology to detect and analyze the copy number variation and gene expression of sheep Y chromosome PRAMEY gene. It mainly includes the following steps: using the DDX3Y gene and β-actin gene that have been verified as single copies by sequencing as references, and using real-time quantitative PCR to identify the copy number and expression level of the PRAMEY gene.

[0033] In order to improve the accuracy of the results, strict experimental operations are carried out according to the requirements of standardized real-time quantitative PCR operations, including: ensuring the extraction of high-quality genomic DNA and RNA, conducting strict target gene primer design...

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Abstract

The invention discloses a method for detecting sheep PRAMEY gene copy number variation and application thereof. The method comprises the following steps: taking whole genome DNA of a ram to be detected as a template, amplifying part of fragments of ram PRAMEY gene and DDX3Y gene through male specific qPCR primer pairs P1 and P2 respectively, serving the multi-copy PRAMEY gene as a detection gene and the single-copy DDX3Y gene as reference genes, and calculating the copy number of the ram PRAMEY gene according to a quantitative result. The method is simple, rapid and convenient to apply and popularize.

Description

technical field [0001] The invention belongs to the field of molecular genetics detection, and in particular relates to a method for detecting the copy number variation of PRAMEY gene (Preferentially expressed antigen in melanoma, Y-linked) of ram Y chromosome. Helicase 3Y-Linked) was used as a reference, and the copy number of the PRAMEY gene in the ram sample was calculated using the method of absolute quantification of the calibration sample. Background technique [0002] With the continuous in-depth study of genome, genome structure variation has been paid more and more attention by scholars. The Y chromosome is a male-specific chromosome and plays an important role in genetic breeding. At least three types of polymorphisms have been found on the Y chromosome: single nucleotide polymorphism and insertion / deletion, short tandem repeat sequence (Short tandem repeaton the Y chromosome, STR) and gene copy number variation (Copy number variation). , CNV). Gene copy number ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6851
CPCC12Q1/6888C12Q1/6851C12Q2600/156C12Q2600/124C12Q2531/113C12Q2537/16C12Q2545/114Y02P60/87
Inventor 乐祥鹏徐海月王丽李发弟李万宏
Owner 甘肃润牧生物工程有限责任公司
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