LncRNA marker related to porcine muscle fiber types and application of lncRNA marker
A technology for muscle fiber types and markers, which is applied to the field of molecular marker preparation in livestock, can solve problems such as reduced pork quality
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Embodiment 1
[0038] Example 1 Different breeds of pig muscle fiber types and high-throughput lncRNA-seq sequencing analysis
[0039] 1) Select Landrace pigs with large differences in carcass muscle fiber types and Guangdong special breed Lantang pigs as research objects; select 3 cases of one-day-old Landrace pigs and three cases of one-day-old Blue Tang pigs to take their back muscle tissue and tissue samples respectively After taking it out, it was quickly frozen in liquid nitrogen for 30 seconds, and then stored in a -80°C refrigerator. Then a part was subjected to paraffin-embedded section and muscle fiber hematoxylin-eosin staining analysis, and the other part was sent to the sequencing company with dry ice for lncRNA sequencing.
[0040] 2) There are obvious differences in muscle fiber appearance in the back muscle tissue of one-day-old lean-meat Landrace pigs and fat-type Blue Tang pigs. The longitudinal sectional area was lower than that of Landrace pigs, and the cross section of a...
Embodiment 2
[0046] Example 2 Fluorescence quantitative PCR detects the expression of LincRNA-ROMT in pig muscle tissue
[0047] Reagent:
[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 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 cases of Landrace pig muscle tissue and 50 cases of Lantang pig muscle tissue, and use the TRIzol method according to the instructions of the Invitrogen kit. The specific steps are as follows:
[0055] 1...
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