where [m.sub.R] is the added reference amount (e.g., 0.011 ng of LSM14B), [N.sub.A] is Avogadro's constant, and MW is the molar weight of the reference DNA (calculated for the plasmid with LSM14B sequence: 2.85 x [10.sup.6] [gmol.sup.-1]).
The selected CpG sites were within "like SM" domain (LSM) family member 14B (LSM14B; cg06096175), zinc finger CCCH-type containing 3 (ZC3H3; cg25834632), and a CpG site not associated with any gene (cg09414987).
Initially, we analyzed serial dilutions of reference DNA (LSM14B) in 2 independent peripheral blood samples (Fig.
Subsequently, we mixed 150 [micro]L of frozen blood (n = 41; validation set III) with our LSM14B reference DNA and analyzed DNAm at the relevant CpG site by pyrosequencing.
 Human Genes: WDR20, WD repeatdomain 20; CD4, CD4 molecule; CD8A, CD8a molecule; FYN, FYN protooncogene, Src family tyrosine kinase; BCL11B, B-cell CLL/lymphoma 11B; WIPI2, WD repeat domain, phosphoinositide interacting 2; SLC15A4, solute carrier family 15 member 4; CENPA, centromere protein A; LSM14B, LSM family member 14B; ZC3H3, zinc finger CCCH-type containing 3.