Fluorescent quantization PCR kit and detection method for detecting cow endometrial cell inflammatory reaction and application of kit

An endometrial and inflammatory reaction technology, applied in the field of PCR amplification, can solve the problems of breeding economic loss, difficulty in in vitro diagnosis of cow's recessive endometritis, etc., and achieves the effects of simple operation and improved economic benefits.

Inactive Publication Date: 2014-09-03
广州中肽天农业科技有限公司
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Problems solved by technology

[0004] Recessive endometritis in dairy cows is a common obstetrical disease after postpartum recovery. Due to the lack of obvious clinical symptoms, in

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  • Fluorescent quantization PCR kit and detection method for detecting cow endometrial cell inflammatory reaction and application of kit
  • Fluorescent quantization PCR kit and detection method for detecting cow endometrial cell inflammatory reaction and application of kit

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Embodiment Construction

[0018] The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores.

[0019] The fluorescent quantitative PCR diagnostic kit for recessive endometritis in cows of the present invention comprises: reverse transcription solution, 2×SYBR GREEN premix solution, primer mixture 1, primer mixture 2, mRNA extract and ultrapure water.

[0020] The components in the kit are as follows:

[0021] Reverse transcription solution: containing DNase (25U / μL), RNase inhibitor (50 U / μL), reverse transcriptase SuperScript Ⅱ (250U / μL).

[0022] 2×SYBR Green master mix: containing Taq deoxyribonucleic acid polymerase (0.1 U / μL), dNTPs substrate (0.4 mmol / L), Mg 2+ (5.0 mmol / L), KCl (100 mmol / L),...

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Abstract

The invention provides a fluorescent quantization PCR kit for detecting cow endometrial cell inflammatory reaction. The fluorescent quantization PCR kit comprises a PCR buffer solution, a TaqDNA polymerase mixture, an SYBRGreen dye, and two pairs of primers, wherein the two pairs of primers include a primer 1 and a primer 2, the primer 1 comprises a forward primer 5'-TATGACCAGGGACCAGGTAC-3' and a reverse primer 5'-CTCTGCCCTTAGGGACTCACAG-3', and the primer 2 comprises a forward primer 5'-ATCGGCAATGAGCGGTTCC-3' and a reverse primer 5'-GTGTTGGCGTAGAGGTCCTTG-3'. The kit disclosed by the invention can be simple and fast, and can relatively and quantitatively detect the transcriptional level of an amyloid protein (SAA) gene in a cow endometrial cell, so as to provide reference for quickly judging whether the dairy cow has recessive endometritis.

Description

technical field [0001] The invention belongs to the technical field of PCR amplification, and in particular relates to a specific fluorescent quantitative PCR kit and a detection method for detecting the inflammatory response of cow endometrium cells. Background technique [0002] Polymerase chain reaction (Polymerase Chain Reaction, PCR) is the most commonly used technique in DNA manipulation technology in vitro, which realizes the exponential amplification of target DNA fragments in the reaction solution. Fluorescence quantitative PCR technology combines real-time fluorescence detection technology and computer analysis technology. Specific fluorescent marker substances are added to the PCR reaction system, and the amplification of DNA can be monitored in real time by detecting the change of fluorescence value in the PCR reaction system after each thermal cycle. and obtain the fluorescence quantitative change curve of each sample, and then obtain the number of PCR cycles (C...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6883C12Q2600/158C12Q2531/113C12Q2563/107
Inventor 张世栋王东升董书伟严作廷王旭荣杨峰
Owner 广州中肽天农业科技有限公司
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