A lncRNA marker related to porcine muscle fiber type and its application
A muscle fiber type and marker technology, applied in the field of livestock molecular marker preparation, can solve the problem of pork quality reduction
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Embodiment 1
[0038] Example 1 Types of porcine muscle fibers of different breeds and high-throughput lncRNA-seq sequencing analysis
[0039] 1) Landrace pigs and Guangdong characteristic breed Lantang pigs with large differences in carcass muscle fiber types were selected as the research objects; 3 one-day-old Landrace pigs and 3 one-day-old Lantang pigs were selected to obtain their back muscle tissue and tissue specimens respectively. After being taken out, it was snap-frozen in liquid nitrogen for 30 seconds, and then stored in a -80°C refrigerator. Then, a part was subjected to paraffin-embedded sections and muscle fiber hematoxylin-eosin staining analysis, and the other part was sent to a sequencing company with dry ice for lncRNA sequencing.
[0040] 2) The apparent differences of muscle fibers in the back muscle tissue of one-day-old lean landrace pigs and fat bluetang pigs were significantly different. The number of muscle fiber nuclei per unit area of Lantang pigs was significant...
Embodiment 2
[0046] Example 2 Fluorescence quantitative PCR detection of LincRNA-ROMT expression in porcine muscle tissue
[0047] Reagents:
[0048] Reverse Transcription Kit: PrimeScript TM RT reagent Kit with gDNA Eraser (Perfect RealTime), #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 application, 120 ℃ high pressure for 20 minutes, and 180 ℃ high temperature drying for more than 2 hours.
[0052] 2) Soak the plastic utensils (eg: EP tube / pipe tip) with 0.1% DEPC water overnight before use, then control the liquid to dry, press 120°C for 20 minutes, and bake it in an oven for use.
[0053] Total RNA extraction:
[0054] 50 cases of Landrace pig muscle tissue and 50 cases of Lantang pig muscle tissue were selected, and the TRIzol method was used according to the instructions of the Invitrogen kit. The specific steps are as follo...
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