Application of C21orf62 gene in liver cancer diagnosis
A gene, liver cancer technology, applied to the C21orf62 gene in the field of liver cancer diagnosis
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Embodiment 1
[0050] Example 1 Screening Gene Markers Related to Liver Cancer
[0051] 1. Sample collection
[0052] 8 cases of normal tissue and liver cancer tissue samples were collected respectively. All the above samples were obtained with the consent of the organizational ethics committee, and the samples were collected and frozen in liquid nitrogen.
[0053] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)
[0054] 1) Sample pretreatment: Take the sample from the -80°C liquid nitrogen tank and put it into a homogenizer soaked in 0.1% DEPC water overnight and sterilized, and grind it into powder; the sample must always be kept in the liquid during the grinding process. In a nitrogen environment, the grinding cup must be pre-cooled, and liquid nitrogen is added while grinding.
[0055] 2) Cleavage: add 1ml Trizol RNA extraction solution to the homogenizer, homogenize at room temperature for 10 minutes, and let stand for 10 minutes to fully lyse the nucle...
Embodiment 2
[0074] Example 2 QPCR sequencing to verify the differential expression of C21orf62 gene
[0075] 1. Large-sample QPCR verification of differential expression of C21orf62 gene. According to the sample collection method in Example 1, 60 cases of liver cancer tissues and 60 normal tissues were selected.
[0076] 2. The specific operation steps of QPCR are as follows:
[0077] (1) RNA extraction
[0078] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:
[0079]1) Add Trizol and place at room temperature for 5 minutes;
[0080] 2) Add 0.2ml of chloroform, vibrate the centrifuge tube vigorously, mix well, and place it at room temperature for 5-10min;
[0081] 3) Centrifuge at 12000rpm for 15min, transfer the upper aqueous phase to another new centrifuge tube (be careful not to absorb the protein material between the two aqueous phases), add an equal volu...
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