Use of AGAP9 in preparation of product for diagnosis of osteoarthritis
A technology for osteoarthritis and products, applied in the field of disease diagnosis
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Embodiment 1
[0045] Example 1 Screening of differentially expressed genes
[0046] 1. Materials:
[0047] Synovial tissues from 5 OA patients (OA group) were obtained from OA patients undergoing knee arthroplasty or synovectomy. The cases used were in line with the diagnostic criteria for OA proposed by Altam. 3 cases of normal synovial tissue (Nor group) came from joint synovial tissue of patients undergoing trauma surgery. All clinical samples used in this study were informed to the patients and approved by the ethics committee of the hospital.
[0048] The clinical information of the selected subjects is shown in Table 1.
[0049] Table 1 Clinical information
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[0051]
[0052] 2. Obtaining tissue RNA
[0053] Total RNA was extracted from tissue samples, and the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of RNA was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. A single library...
Embodiment 2
[0071] Example 2 Large sample verification screened out differentially expressed genes
[0072] Based on the results of previous sequencing and according to the size of the P value, we selected the AGAP9 gene for verification.
[0073] 1. Sample collection
[0074] According to the method of Example 1, 42 cases of osteoarthritis synovial tissue and 42 normal synovial tissue were collected.
[0075] 2. Validation at the mRNA level
[0076] 2.1 Extract tissue RNA
[0077] Step is with embodiment 1.
[0078] 2.2 Reverse transcription
[0079] Reverse transcription using Primescript 1 st strand cDNA synthesis kit kit, the operation steps are as follows:
[0080] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 2:
[0081] Table 2 Reaction liquid
[0082]
[0083]
[0084] (2) Incubate at 70°C for 5 minutes, then rapidly cool to 4°C;
[0085] Add the following reaction reagents into the microcentrifuge tube to make a reaction syst...
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