Application of long-chain non-coding RNA LINC00263 in preparation of adenocarcinoma-of-lung prognosis detection preparation
A long-chain non-coding, lung adenocarcinoma technology, applied in the field of tumor molecular biology, can solve the problems of no reference standard for the prognosis of patients with lung adenocarcinoma, inability to adapt to the prognosis of patients with lung adenocarcinoma, and no specific indicators, etc. To achieve the effect of easy promotion and application, far-reaching clinical significance and promotion, and cost-effective effect
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Embodiment 1
[0020] Example 1 Preparation of long-chain non-coding RNA LINC00263 for prognostic detection kit of lung adenocarcinoma (50 reactions)
[0021] 1. RNA stabilization solution 50ml
[0022] 2. Isopropanol 100ml
[0023] 3. Trichloromethane 100ml
[0024] 4.Trizol 50ml
[0025] 5. 10ml of enzyme-free water
[0026] 6. 1μM random reverse transcription primer 50ul
[0027] 7. 5×Reverse Transcription Buffer 200ml
[0028] 8. 10mM triphosphate base deoxynucleotide 100ul
[0029] 9. 40U / μl RNase inhibitor 500ul
[0030] 10. 200U / μl MMLV reverse transcriptase 50ul
[0031] 11.Premix Ex Taq 50ul
[0032] 12. 10μM long-chain non-coding RNA LINC00263 real-time fluorescent quantitative PCR specific primers:
[0033] Forward primer 5'TCGGATAGGAGTGTCAGG-3'(SEQ ID NO.1),
[0034] Reverse primer 5'TTCAGTTGCTTCAGGTCAT-3'(SEQ ID NO.2),
[0035] 13.10μM ACTB real-time fluorescence quantitative PCR specific primers:
[0036] The forward primer is 5'-CACCATTGGCAATGAGCGGTTC-3' (SEQ ID NO.3),
[0037] The reverse primer i...
Embodiment 2
[0038] Example 2 Detection of long-chain non-coding RNA LINC00263 in lung adenocarcinoma tissue
[0039] 1. Preservation of lung adenocarcinoma tissue: Collect the lung adenocarcinoma tissue to be tested and store it in a cryopreservation tube containing RNA stabilizing solution, and put it in a refrigerator at -80℃ for later use.
[0040] 2. Extraction of RNA from tissues: Take an appropriate amount of specimens and add liquid nitrogen to grind the specimens in a mortar baked at 180°C for 6-8 hours, grind them to powder form, and then add 1ml Trizol mortar specimens into the mortar and grind them into After the liquid state, transfer to the centrifuge tube, add 200μl / ml of chloroform to the centrifuge tube, shake by hand for 15-30s, place on ice for 5 minutes, centrifuge at 12000g at 4℃ for 15 minutes; carefully take the upper water phase into a new centrifuge tube and add Mix well with 0.5ml of pre-cooled isopropanol, stand in the refrigerator at -20°C for 20min, centrifuge at 12...
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