DNA library was prepared using the VariantPro™ Mitochondrion Panel Library Preparation Kit (LC Sciences). The presence of the desired fragments and the purity of the indexed libraries were analysed on Agilent 2100 Bioanalyzer (Agilent Technologies) using High-Sensitivity DNA Analysis Kit (Agilent Technologies). The concentrations of the libraries were measured using Qubit Fluorometer (Thermo Fisher Scientific). Equimolar amounts of the 45 indexed libraries were pooled to obtain a 4 nM library mixture. After denaturing, and further diluting, the final 10 pM of library mixture was loaded into Illumina cartridge. Sequencing was performed using the Illumina MiSeq Reagent v2 kit (500 cycles) on the Illumina MiSeq instrument following the manufacturer’s instructions (Illumina).
Variantpro mitochondrion panel library preparation kit
Manufactured by LC Sciences
The VariantPro™ Mitochondrion Panel Library Preparation Kit is a laboratory equipment product designed for the preparation of mitochondrial DNA libraries. It provides the necessary reagents and protocols for the amplification and sequencing of targeted mitochondrial regions.
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Lab products found in correlation
High sensitivity dna analysis kit, by Agilent Technologies (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Miseq reagent kit v2, by Illumina (1 mentions)
Qubit fluorometer, by Thermo Fisher Scientific (1 mentions)
Hiseq x ten platform, by Illumina (1 mentions)
Agencourt ampure xp bead, by Beckman Coulter (1 mentions)
2 protocols using variantpro mitochondrion panel library preparation kit
1
Mitochondrial DNA library preparation and sequencing
2
Mitochondrial DNA Somatic Mutation Analysis
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In 68 out of the 84 available samples, mtDNA somatic mutation analysis was performed by LC Sciences (Hangzhou, China) using VariantPro Mitochondrial Panel. Briefly, mtDNA-targeted libraries were prepared using the VariantPro™ Mitochondrion Panel Library Preparation Kit (LC Sciences). The PCR product was purified by Agen-court AMPure XP beads (Beckman Coulter Genomics, UK), and diluted to 20 pmol/L, then directly sequenced on the Illumina Hiseq X Ten platform based on the 150-bp paired-end reads. Raw reads were processed following the Genome Analysis Toolkit best practices guideline. Reads were trimmed using Trim Galore and cutadapt40 (link) to remove adapters and bases. The trimmed reads were then aligned to the human mtDNA relative to the revised Cambridge Reference Sequence (rCRS,41 (link) GeneBank accession NC012920.1) using Burrows-Wheeler Alignment.42 (link)
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