Application of long-chain non-coding RNA in tumors
A reagent and technology for rectal adenocarcinoma, applied in the field of biomedicine, can solve the problem that the function of long non-coding RNA needs to be further elucidated
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Embodiment 1
[0056] Example 1 Screening for gene markers associated with rectal adenocarcinoma
[0057] 1. Sample collection
[0058] Tissue samples of 5 cases of rectal adenocarcinoma and corresponding paracancerous tissue samples were collected. All cases had not received chemotherapy and radiotherapy before surgery, and other neoplastic diseases, autoimmune diseases and severe chronic diseases were excluded. All patients had Informed consent was obtained and approval was obtained from the organizational ethics committee.
[0059] 2. Preparation of RNA samples
[0060] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.
[0061] 3. Total RNA quantification and purity analysis
[0062] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of the RNA was detected by agarose gel electrophoresi...
Embodiment 2
[0071] Example 2 QPCR sequencing to verify the differential expression of the LOC105378728 gene
[0072] 1. Large-sample QPCR verification of differential expression of LOC105378728 gene. According to the sample collection method in Example 1, 30 cases of rectal adenocarcinoma paracancerous tissues and 30 cases of rectal adenocarcinoma tissues were selected.
[0073] 2. RNA extraction
[0074] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.
[0075] 3. QPCR
[0076] 1) Reverse transcription reaction
[0077] Use Tiangen’s FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) for lncRNA reverse transcription, first remove the genomic DNA reaction, add 5×gDNA Bμffer 2.0μl, total RNA 1μg, add RNase Free ddH 2 O Make the total volume to 10μl, heat in a water bath at 42°C for 3min, then add 10×Fast RT Bμffer 2.0μl, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O 5.0 μl, after m...
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