Molecular marker for diagnosis of lung adenocarcinoma and application thereof
A lung adenocarcinoma, material technology, applied in the field of lung adenocarcinoma diagnosis and molecular markers, can solve the problems of expression and function slowly unfolding
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Embodiment 1
[0071] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma
[0072] 1. Sample collection
[0073] A total of 35 lung adenocarcinoma tissues and corresponding paracancerous tissues were collected. All patients had not received other treatments before surgery. Among them, 4 lung adenocarcinoma tissues and corresponding paracancerous tissues were selected for high-throughput sequencing.
[0074] 2. RNA sample preparation and quantitative analysis
[0075] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.
[0076] 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 electrophoresis, and the RIN value was determined by Agilent2100. A single library construction requires a total amount of RNA of 5 μg, a concentration ≥ 200 ng / μL,...
Embodiment 2
[0094] Example 2 QPCR sequencing to verify differential expression of RP11-492E3.2 gene
[0095] 1. The differential expression of RP11-492E3.2 gene was verified on 35 collected tissue samples.
[0096] 2. RNA extraction
[0097] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.
[0098] 3. QPCR
[0099] 1) Reverse transcription reaction
[0100] Use the FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) from Tiangen Company to carry out lncRNA reverse transcription, first remove the genomic DNA reaction, add 5×gDNA Bμffer 2.0μl, total RNA 1μg, add RnaseFree ddH 2 O to bring the total volume to 10 μl, and heat in a water bath at 42°C for 3 min.
[0101] Mix 10× Fast RT Bμffer 2.0μl, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O5.0 μl, after mixing, add to the above-mentioned test tube and mix together a total of 20 μl, heat at 42°C for 15 minutes in a water bath, and he...
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