Molecular marker for cancer diagnosis
A technology of gastric adenocarcinoma and reagents, applied in the field of molecular markers for cancer diagnosis, can solve problems such as unclear master-slave relationship of lncRNA and unclear role of 1ncRNA
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Embodiment 1
[0036] Example 1 Screening for gene markers associated with gastric adenocarcinoma
[0037] 1. Sample collection
[0038] The cancer tissues and corresponding paracancerous tissue samples of 4 cases of gastric adenocarcinoma were collected for high-throughput sequencing. All patients did not receive chemotherapy, radiotherapy and endocrine therapy before operation.
[0039] 2. RNA sample preparation and quality analysis
[0040] Total RNA was extracted using Tiangen Animal Tissue Total RNA Extraction Kit (catalogue number: DP431), and the steps are detailed in the instructions.
[0041] 1) Homogenization treatment
[0042] Add 300μl Lysis Solution RL for every 10-20mg tissue, and grind the tissue thoroughly with a grinding pestle; then add 590μl RNase-Free ddH to the homogenate 2 O and 10 μl Proteinase K, mix well and treat at 56°C for 10-20min.
[0043] 2) Centrifuge at 12,000rpm for 2-5min, and take the supernatant for the following operations.
[0044] 3) Slowly add 0....
Embodiment 2
[0076] Example 2QPCR sequencing verification of differential expression of RP11-576I22.2 gene
[0077] 1. The differential expression of the RP11-576I22.2 gene was verified by large-scale QPCR on the cancer tissue samples and para-cancerous tissue samples collected from 31 gastric adenocarcinoma patients according to the collection method in Example 1.
[0078] 2. RNA extraction
[0079] The total RNA was extracted using Tiangen Animal Tissue Total RNA Extraction Kit (Cat. No. DP431). See Example 1 for specific steps.
[0080] 3. QPCR
[0081] Primers were designed according to the gene sequences of RP11-576I22.2 and GADPH, and the primer sequences were as follows:
[0082] RP11-576I22.2:
[0083] Forward primer: 5′-AGTATGAGGTAGCCAACA-3′ (SEQ ID NO.1)
[0084] Reverse primer: 5'-AACTTGACTTCTGACATCTT-3' (SEQ ID NO.2)
[0085] GAPDH:
[0086] Forward primer: 5'-AATCCCATCACCATCTTCCAG-3' (SEQ ID NO.3)
[0087] Reverse primer: 5'-GAGCCCCAGCCTTTCTCAT-3' (SEQ ID NO.4)
[0088...
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