Application of ENSG00000267549 in judgment of metastasis of osteosarcoma
A technology for osteosarcoma and products, applied in the field of biomedicine, can solve problems such as no specific model
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Embodiment 1
[0041] Example 1 Screening of lncRNAs associated with osteosarcoma
[0042] 1. Sample collection and clinical information
[0043] The tumor tissues of 3 patients with osteosarcoma metastases (metastatic lesions, the metastatic site is the lung) and 3 patients with non-metastatic osteosarcoma (primary lesions) were collected. The clinical information of the patients is shown in Table 1. The collection of all samples was approved by the ethics committee, and the patients gave informed consent.
[0044] Table 1 Clinical information of patients
[0045]
[0046] 2. Preparation of RNA samples
[0047] RNA samples were extracted using the tissue RNA extraction kit from Invitrogen, see the instruction manual for details.
[0048] 3. Quality analysis of RNA samples
[0049] Nanodrop2000 was used to detect the concentration and purity of the extracted RNA, agarose gel electrophoresis was used to detect RNA integrity, and Agilent2100 was used to determine the RIN value. Concent...
Embodiment 2
[0056] Example 2 QPCR verification of differentially expressed ENSG00000267549
[0057] 1. According to the high-throughput sequencing results, ENSG00000267549 was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 36 patients with primary tumor and 33 patients with tumor metastasis were selected.
[0058] 2. RNA extraction
[0059] RNA samples were extracted using the tissue RNA extraction kit from Invitrogen, see the instruction manual for details.
[0060] 3. Reverse transcription:
[0061] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl RNase inhibitor and RNase free water , the final volume was 20 μl, and incubated at 37°C for 1h.
[0062] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.
[0063] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers...
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