IncRNA related to chicken sexual maturity or heterosis, target gene regulated by lncRNA and application of lncRNA
A hybrid vigor, mature technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of LncRNA without literature reports
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[0055] Example 1 Screening of molecular markers related to chicken sexual maturity
[0056] ⒈ Test method
[0057] ⑴ group building
[0058] Choose White Leghorn Chicken (WW) and Beijing Oil Chicken (YY), and perform pure breeding and positive and negative crosses according to the ratio of male to female to form 2 pure breeding combinations (WW, YY) and 2 hybrid combinations (WY, YW) ). The chickens are raised in the same house with the same management method.
[0059] (2) Phenotypic analysis and sample collection
[0060] When the chickens were 22 weeks old, 6 individuals were selected from the WW, YY, WY, and YW combinations according to the individual pubic width for ovarian tissue sample collection and fallopian tube length measurement, and the ovarian tissue was put into liquid nitrogen for preservation, and then transferred to - 80 ℃ refrigerator for spare use, the entire sampling process to maintain a sterile environment. At the same time, the blood of the selected...
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[0063] Example 2 LncRNA and its target gene regulatory relationship and associated signaling pathway analysis
[0064] ⒈ Library sequencing
[0065] Sample rRNA was removed using the Epicentre Ribo-ZeroTM kit. Fragmentation is then performed, and first-strand cDNA is synthesized using random primers and reverse transcriptase, and then double-strand cDNA is synthesized. The purified double-stranded cDNA is then subjected to end repair, A is added, and a sequencing adapter is connected, and finally a cDNA library is obtained by PCR amplification. After passing the quality inspection, the library was sequenced using an Illumina HiSeq sequencer to obtain the original sequencing data.
[0066] ⒉ LncRNA Screening
[0067] Clean reads were obtained from raw sequencing data after quality control. Taking the Gallus-gallus version 6.0 genome as the reference genome, and aligning and assembling transcripts according to the Hisat2-Stringtie process, LncRNA prediction was performed. F...
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