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Specific primer combination used for detecting gene expression of fine-wool sheep skin follicles

A technology for skin hair follicles and gene expression, which is applied in the field of biological genes, can solve the problems of complicated operation, low accuracy, and the need for semi-quantitative reverse transcription PCR, and achieves the effects of high sensitivity and simple operation.

Inactive Publication Date: 2017-10-20
QINGDAO AGRI UNIV
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  • Claims
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Problems solved by technology

The current methods that use protein as the detection target are usually cumbersome and time-consuming
Semi-quantitative reverse transcription PCR requires electrophoresis detection, and the accuracy is not high enough
[0004] To sum up, the problems existing in the prior art are: the current methods that use protein as the detection target are usually cumbersome and time-consuming.
Semi-quantitative reverse transcription PCR requires electrophoresis detection, and the accuracy is not high enough

Method used

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  • Specific primer combination used for detecting gene expression of fine-wool sheep skin follicles
  • Specific primer combination used for detecting gene expression of fine-wool sheep skin follicles
  • Specific primer combination used for detecting gene expression of fine-wool sheep skin follicles

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Effect test

Embodiment 1

[0061] Relative expression of EphrinA3 in the flank skin of fine-wool sheep in different periods

[0062] After extracting total skin RNA, it was detected by effendorf Biophotometer Plus spectrophotometer, OD260 / 280 was 1.8-2.0, and detected by 1.0% agarose gel electrophoresis, both 28sRNA and 18sRNA were bright, and the ratio of the two bands of 28s and 18s was close 2:1. The results showed that the integrity of the extracted total RNA was good, and the concentration and purity could meet the requirements of subsequent experiments.

[0063] There is only one obvious peak in the melting curve peak diagram of Ephrin A3 and the internal reference gene GAPDH, indicating that the fluorescent signals are all from specific amplification products in the process of real-time fluorescent quantitative PCR, and neither Ephrin A3 nor the internal reference gene GAPDH produces non-specific amplification And primer dimers, the data is reliable, and the next step of data analysis can be per...

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Abstract

The invention belongs to the field of biological genes, and discloses a specific primer combination used for detecting the gene expression of fine-wool sheep skin follicles. The specific primer combination comprises a specific primer of sheep EphrinA3 gene and a specific primer of GAPDH gene; and the nucleotide sequence of the specific primer of the sheep EphrinA3 gene is represented by SEQ ID NO.1, the nucleotide sequence of the specific primer of the GAPDH gene is represented by SEQ ID NO.2, and the designed primers have no specific amplification. A fluorescent quantitative PCR method used in the invention is a high-sensitivity simple and quick detection method, is simple to operate, and can be used to SYBR green dye method detection without using a fluorescent probe; and a fluorescent quantitative PCR relative quantification method is used to compare with housekeeping gene having a stable expression level in order to reflect the expression levels of the target genes, so the study of the EphrinA3 expression level can be well assisted.

Description

technical field [0001] The invention belongs to the field of biological genes, and in particular relates to a combination of specific primers for detecting gene expression in skin hair follicles of fine-wool sheep. Background technique [0002] Ephrin is a ligand member of the Eph receptor family and is mainly expressed on the cell surface. Ephrin ligands have 8 members, which are divided into two subfamilies, EphrinA and EphrinB, according to their attachment methods to the cell membrane. Among them, the EphrinA subfamily has 5 members, namely EphrinA1-A5. [0003] Suo et al. found that feeding mice high levels of dietary vitamin A can enhance Wnt signaling to activate hair follicle stem cells. Overexpression of Wnt in injured skin of adult mice can increase the number of regenerated hair follicles, and the Wnt receptor Frizzled 1 or 2 Activation stimulates the expression of Ephrin A3. Yamada et al. also found that the expression of EphrinA3 in the dermal papilla cells of...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6806C12Q1/6888C12Q2531/113
Inventor 贺建宁王春亮柳楠刘积凤
Owner QINGDAO AGRI UNIV
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