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Table 1 Overview of genome-wide 5hmC quantification methods

From: Towards precision medicine: advances in 5-hydroxymethylcytosine cancer biomarker discovery in liquid biopsy




Whole-genome TAB-Seq [70] (direct measurement of 5hmC)


Single nucleotide resolution

Absolute methylation value

High sequencing depth required

Input > 100 ng DNA

Whole-genome oxBS-Seq [71, 74] (inference of 5hmC)

RRBS TAB-Seq [91] (direct measurement of 5hmC)

Single nucleotide resolution

Absolute methylation value

Targets CpG islands

Input > 500 ng DNA

RRBS oxBS-Seq [52] (inference of 5hmC)

~ 60 million PE reads/sample

Input > 500 ng DNA

Nano-hmC-Seal [53] (direct measurement of 5hmC)

Input 5 ng DNA

Excellent genomic coverage

High specificity for 5hmC

~ 30 million PE reads/sample

Lower resolution

  1. 5hmC, 5-hydroxymethlcytosine; TAB-Seq, Tet-assisted bisulfite sequencing; oxBS-Seq, oxidative bisulfite sequencing; PE, paired-end