Fluorescence quantitative PCR detection primer and method for type and composition of yak muscle fibers
A technique for muscle fiber type and fluorescence quantification, which is applied to the detection primers and detection fields of fluorescence quantitative PCR composed of yak muscle fiber types, and achieves the effects of low cost, high efficiency and small workload.
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[0040] 1. Sample collection:
[0041] The longissimus dorsi muscle tissue of yaks was collected quickly after slaughter and stored in liquid nitrogen.
[0042] 2. Reagents:
[0043] RNAsimple total RNA extraction kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.;
[0044] PrimeScript? RTreagentKitwithgDNAEraser: Takara Corporation;
[0045] SYBR?PremixExTaq?II (PerfectRealTime): Takara Company;
[0046] Primer synthesis: American life company.
[0047] 3. Main instruments and equipment:
[0048] PCR instrument: Bio-rad company;
[0049] CFX-96 fluorescent quantitative PCR instrument: Bio-rad company;
[0050] Nanodrop2000 Ultra Micro Spectrophotometer: ThermoScientific Company.
[0051] 4. Detection method:
[0052] a) Design and synthesis of primers:
[0053] According to the cDNA sequence differences of different subtypes of yak MyHC gene, primers were designed using Primer Premier5.0 and Oligo6.0 software. The company synthesizes TaqMan probes. The sequences ...
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