The miRNA markers related to pig carcass lean meat percentage and its application
A technology of markers and lean meat percentage, applied in the field of miRNA markers
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Embodiment 1
[0037] Example 1 High-throughput miRNA-seq identification of differentially expressed miRNAs in different breeds of pig muscle tissue
[0038] 1) Landrace pigs with large differences in carcass lean meat percentage and Lantang pigs, a special breed of Guangdong, were selected as the research objects; 3 cases of Landrace pigs and 3 cases of Lantang pigs were selected to obtain their back muscle tissues, and the tissue samples were quickly frozen after they were taken out. In liquid nitrogen for 30 seconds, then stored in a -80°C refrigerator, and then sent to a sequencing company with dry ice for small RNA sequencing.
[0039] 2) Use FastQC (http: / / www.bioinformatics.babraham.ac.uk / projects / fastqc / ) software to evaluate the overall quality of sequencing data, including base quality value distribution, quality value position distribution, GC content, PCR duplication content, etc.
[0040] After high-throughput sequencing, the total number of reads obtained from the three back m...
Embodiment 2
[0044] Example 2 Detection of ssc-miR-423-5p expression in porcine muscle tissue by fluorescent quantitative PCR
[0045] Reagent:
[0046] Reverse transcription kit: PrimeScript TM RT reagent Kit with gDNA Eraser (Perfect RealTime), #RR047Q, Takara;
[0047] Quantitative PCR reagents: Fast qPCR Mix, #RR430A, Takara.
[0048] Relevant experimental items to RNase treatment:
[0049] 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.
[0050] 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.
[0051] Total RNA extraction:
[0052] 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:
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