A lncRNA marker and its application in the diagnosis of acute myocardial infarction
A kind of acute myocardial infarction, primer pair technology, applied in the field of biomedicine
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Embodiment 1
[0027] 1. Processing of plasma samples
[0028] The collected blood was refrigerated at 4°C for 30 minutes, then centrifuged at 3000 rpm for 15 minutes at room temperature, and the upper layer of plasma was transferred to a 1.5ml centrifuge tube and stored in a -20°C refrigerator for later use.
[0029] 2. Extraction of plasma total RNA
[0030] (1) Thaw the separated plasma on ice, take 250μl plasma sample into a 2ml centrifuge tube, add 750μl Trizol, mix well by pipetting, and let stand for 5min;
[0031] (2) Add 200 μL of chloroform, mix upside down 20 times, let stand at room temperature for 5 minutes, centrifuge at 12000 rpm for 15 minutes;
[0032] (3) Carefully transfer the upper aqueous phase to a new RNase-free centrifuge tube, add an equal volume of isopropanol, mix well and place on ice for 10 minutes;
[0033] (4) Centrifuge at 13,000 rpm, 4°C for 10 minutes, discard the supernatant, and keep the precipitate;
[0034] (5) Add 500 μL pre-cooled 70% ethanol to dis...
Embodiment 2
[0062] A fluorescent quantitative PCR kit for diagnosing acute myocardial infarction
[0063] 1. Composition
[0064] LINC01104 primer pair, GAPDH primer pair, SYBR Green fluorescent dye, and ddH2O.
[0065] LINC01104
[0066] Forward primer 5'-AGGGCATGACAGGCAATTCA-3'
[0067] Reverse primer 5'-AGTGATCCTCTTGCCTTGGC-3'
[0068] GAPDH
[0069] Forward primer 5'-ACGTGTCAGTGGTGGACCTG-3'
[0070] Reverse primer 5'-GTGTAGCCCAGGATGCCCTT-3'
[0071] 2. How to add
[0072]
[0073] 2. Reaction conditions
[0074] Reaction conditions: 95°C for 10min; 95°C for 15s, 60°C for 60s for 40 cycles.
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