Amplicon sequencing of 3P pig: liver (left)
Two target mtDNA amplicons, size 338bp and 262bp, were amplified separately using primer Set1 (F=GCCTATTCTGAATACTAGTAGC, R= GTTTGCATACGAATGGAATGC) and Set4 (F=TACTAGTGTTTGCAGCCTGC, R=GGAAAAATTAAGGCTGTTTTCGC), respectively. The amplicons were pooled and library preparation using Nextera DNA Flex Library Preparation Kit (Illumina). The library were 150 bp paired-end sequenced using NovaSeq 6000 SP Reagent Kit (Illumina). Image analysis was performed in real-time by the NovaSeq Control Software (NCS) v1.7.5, and Real-Time Analysis (RTA) v3.4.4 performed real-time base calling on the NovaSeq instrument computer. Then the Illumina bcl2fastq2.20.0.422 pipeline was used to generate the sequence data.