Molecular marker related to porcine intermuscular fat and application of molecular marker
An intermuscular fat and molecular marker technology, applied in the field of livestock molecular marker preparation, can solve the problems of difficult separation of intermuscular fat, difficulty in specific research and evaluation, etc., and achieve the effect of high specificity and accuracy
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Embodiment 1
[0039] Example 1 Based on 10×Genomics single-cell RNA sequencing technology, the specific expression gene SNCA of porcine intermuscular adipocytes was obtained
[0040] Due to the difficulty in anatomically separating the intermuscular adipose tissue of livestock and poultry, ordinary transcriptome sequencing technology cannot achieve specific analysis of porcine intermuscular adipose tissue. However, 10×Genomics single-cell RNA sequencing technology is based on the Gemcode microfluidic platform, which can Single intermuscular adipocytes and gel beads containing 10× barcode information are wrapped in oil droplets to form a "water-in-oil" structure (GEMs). In GEMs, the cells are cleaved to release mRNA, which is generated by reverse transcription for sequencing cDNA, through high-throughput sequencing to obtain the gene expression of a single cell.
[0041]1) The lean-meat Landrace pigs with large differences in carcass meat quality and the fat-type Lantang pigs of Guangdong sp...
Embodiment 2
[0047] Example 2 Fluorescence quantitative PCR technique verification SNCA gene is used as the molecular marker reagent of porcine intermuscular fat:
[0048] Reverse transcription kit: PrimeScript TM RT reagent Kit with gDNA Eraser (PerfectReal Time), #RR047Q, Takara;
[0049] Quantitative PCR reagents: Fast qPCR Mix, #RR430A, Takara.
[0050] Relevant experimental items to RNase treatment:
[0051] 1) Rinse and soak all glassware with DEPC before use, high pressure at 120°C for 20 minutes, and dry at high temperature at 180°C for more than 2 hours.
[0052] 2) Soak plastic utensils (such as EP tubes / tips) in 0.1% DEPC water overnight before use, and then dry the liquid, 120°C and high pressure for 20 minutes, and dry in an oven for later use.
[0053] Total RNA extraction:
[0054] Select 50 lean-meat Landrace pig muscle tissues and 50 fat-type Blue Tang pig muscle tissues, and use the TRIzol method according to the instructions of the Invitrogen kit. The specific ste...
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