Biomarker long-chain non-coding RNA LINC01728 and kit for predicting prognosis of colon cancer
A biomarker and long-chain non-coding technology, which is applied in the field of biomarker long-chain non-coding RNALINC01728 and kits, can solve problems that have not been reported, improve survival rate, improve postoperative quality of life, and have far-reaching clinical significance Effect
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Embodiment 1
[0018] The biomarker used to predict the prognosis of colon cancer is long-chain non-coding RNA LINC01728, and the nucleic acid sequence of the long-chain non-coding RNA LINC01728 is shown in SEQ NO:1.
[0019] The preparation for predicting the prognosis of colon cancer is a real-time fluorescent quantitative PCR detection kit, a real-time fluorescent quantitative PCR detection kit, including specific primers for detecting the expression of the target gene LINC01728, the primer sequences are SEQ NO:2 and SEQNO:3, real-time fluorescent The quantitative PCR detection kit also includes specific primers for the expression of the internal reference gene TUBA1A, and the primer sequences are SEQ NO:4 and SEQ NO:5.
[0020] Preparation of reagents for detecting the expression of LINC01728 is used to prepare a kit for the prognosis of patients with colon cancer (for 50 reactions), and the required reagents include: SYBR Green qPCR Mix 500 μL, 3 μM target gene LINC01728-specific primer ...
Embodiment 2
[0024] Detection of LINC01728 in Tissue Samples
[0025] 1. Collect the colon cancer tumor tissue or normal control tissue to be tested, wash it with normal saline, put it into a cryopreservation tube filled with a solvent that inhibits RNA degradation, and put it in a -80°C refrigerator for later use.
[0026] 2. Extraction of RNA from tissues
[0027] (1) Add liquid nitrogen to the mortar first, then cut the tissue into small pieces and grind it into powder in liquid nitrogen, take 100mg of tissue powder with a liquid nitrogen pre-cooled spoon and add it to the EP tube filled with 1ml of Trizol solution middle. The total volume of tissue powder should not exceed 10% of the volume of Trizol used, and it should be thoroughly mixed evenly.
[0028] (2) Leave at room temperature for 5 minutes, then add 200 μL of chloroform, tightly cap the EP tube and shake vigorously for 0.5 minutes. Centrifuge at 12,000 rpm for 10 minutes.
[0029] (3) Take the upper aqueous phase into a n...
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