Decontamination effectiveness.

<p>To measure decontamination potential we mixed equimolar 5me-dCTP amplified mtDNA into non-methylated Target mtDNA. Methylated and non-methylated DNA were from mtDNA haplogroups differing in 8 loci. Each haplogroup mtDNA sample was barcoded with unique DNA barcodes prior to pooling, decontamination and amplification. Complete decontamination was measured via sequencing the mixed libraries to 10,000× coverage and measuring heteroplasmy levels with and without MspJI decontamination. MspJI digestion removed 100% of expected heteroplasmy contaminants(red) suggesting it can decontaminate up to equimolar contamination events. Undigested pooled libraries were sequenced as a control (blue) and exhibited 35–65% heteroplasmy levels. These artificial heteroplasmies were produced by pooling a methylated mitochondrial Long Range PCR product from a different haplogroup into a non methylated product. This haplogroup is completely removed by the decontamination methods described.