Ultra-early detection kit for serum miRNAs in patients with alcoholic femoral head necrosis
A femoral head necrosis, alcohol-based technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., to achieve good clinical application prospects and good accuracy
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Embodiment 1
[0034] (1) RT-PCR primer synthesis:
[0035] Synthesize miR-127-3p, miR-1, miR-628-3p, miR-432-5p, miR-483-3p, miR-483-5p, miR-885-5p, miR-423-5p according to the following table ( The sequences of RT-PCR detection primers are shown in Table 1.
[0036] Table 1 is the sequence of RT-PCR detection primer GSP and primer R of each miR
[0037]
[0038]
[0039] (2) Detection of miRs expression level
[0040] 1) Synthesis of cDNA template by reverse transcription
[0041] RNA was extracted from the serum or plasma of AIONFH patients and non-AIONFH patients, and reverse-transcribed into cDNA templates. The reaction system for cDNA synthesis by reverse transcription is as follows:
[0042]
[0043] The prepared cDNA template, nuclease free water and SYBR TM Put the Green master mix on ice to dissolve for 15-20 minutes. SYBR TM Green master mix should be dissolved in the dark, and should be inverted before use, and other reagents should be shaken to mix. All reagents...
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