MiRNA marker in serum exosome related to early diagnosis of lung adenocarcinoma and application
A technology of exosomes and markers, applied in the fields of biotechnology and diagnostics, to achieve the effect of convenient diagnosis and improved clinical treatment effect
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[0026] Serum exosome extraction:
[0027] 5ml of cubital venous blood was collected with a coagulation-promoting tube, centrifuged at 3000rpm for 10min, and the supernatant serum was absorbed. The serum was centrifuged at 4°C, 20000g for 30min, and the cell debris was discarded. The obtained serum crude extract was filtered through a 0.22um filter membrane, added with PBS, and centrifuged at 4°C and 150000g for 90min. After centrifugation, discard the PBS, add PBS to suspend and wash the precipitate, and centrifuge at 4°C and 150,000g for 60min. Serum exosomes were obtained, and 500ul Trizol was added to extract RNA.
[0028] Sample total RNA extraction:
[0029](1) Before the experiment, wipe the experimental bench with absolute ethanol to reduce the dust on the experimental bench and disinfect it; (2) Take out the sample and thaw it; (3) After centrifuging the lysate containing Trizol LS at 12000rmp for 10min at 4°C, absorb the supernatant To an RNase-free centrifuge tub...
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