A diagnostic biomarker for colorectal cancer and its application
A biomarker, colorectal cancer technology, applied in biochemical equipment and methods, microbial determination/inspection, organic compound library, etc. High requirements and other problems, to achieve the effect of improving the 5-year survival rate, broad application prospects, and timely diagnosis
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Embodiment 1
[0053] Example 1: High-throughput chip processing and analysis of circular RNA
[0054] Firstly, the raw data is filtered, and then the data with low-quality signal and noise pollution are removed, and the effective circular RNA expression value is obtained after normalization. On the basis of the above analysis, a series of statistical and visual analysis can be carried out.
[0055] In this study, the multiple of change (Fold Change, FC) ≥ 1.5 and P figure 2 ). Then the screened differentially expressed mRNAs were represented by a heat map ( image 3 ).
Embodiment 2
[0056] Example 2: Detection of differential expression of hsa_circ_0006110 gene in colorectal cancer cell lines and normal intestinal epithelial cell lines by fluorescent quantitative PCR
[0057] 1. Cultivate human colorectal cancer cell lines (SW620, HCT116 and RKO) and human normal intestinal epithelial cell line NCM460. Each 100U / mL of DMEM medium at 37°C and 5% CO 2 Cultured in an incubator, the culture medium was changed every 2-3 days and subcultured. Human intestinal cancer HCT116, SW620 and RKO cell lines were treated with DMEM (containing 10% fetal bovine serum) at 37°C and 5% CO 2 cultured in an incubator.
[0058] 2. When the confluence of the cells in the 60mm culture dish reaches 80%, discard the culture medium, add 1mL sterile PBS to wash, discard the PBS and repeat once, add 1mL Trizol RNA extraction solution, and gently shake the culture dish left and right , so that all the Trizol RNA extraction solution was submerged at the bottom of the culture dish, gen...
Embodiment 3
[0076] Example 3: RT-PCR reaction detection of differential expression of hsa_circ_0006110 gene in colorectal cancer and paracancerous tissues.
[0077] 1. Collect 25 pairs of colorectal cancer tissue and corresponding distant normal intestinal tissue samples. Each patient sample has a clear diagnosis record, and with the consent of the ethics committee, the samples are collected and frozen in liquid nitrogen.
[0078] 2. Preparation of RNA samples
[0079] (1) Pretreatment of the sample: Take the sample out of the liquid nitrogen tank, put the sample into a 2mL sterile EP tube, add 1mL Trizol RNA extraction solution, soak it in 0.1% DEPC water overnight and cut it into pieces with sterilized scissors. Grind thoroughly in a tissue homogenizer.
[0080] 3. RNA extraction, reverse transcription, fluorescent quantitative PCR and data analysis methods are the same as in Example 2.
[0081] 4. Results: The expression level of hsa_circ_0006110 was detected in 25 pairs of colorecta...
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