Libraries are prepared using as little as 10 ng input DNA and the supplied NEBNext Ultra II reagents and the optimized EM-seq Adaptor. TET2 then oxidizes 5-mC and 5-hmC, providing protection from deamination by APOBEC in the next step. In contrast, unmodified cytosines are deaminated to uracils. Libraries are then amplified using a NEBNext master mix formulation of Q5U® (a modified version of Q5® High-Fidelity DNA Polymerase), and sequenced using Illumina instrumentation.

The consistently high conversion performance and minimized DNA damage with the EM-seq protocol, in combination with highly efficient Ultra II library prep, result in superior detection of CpGs with fewer sequencing reads.

Features:
·Superior sensitivity of detection of 5-mC and 5-hmC.
·Greater mapping efficiency.
·More uniform GC coverage.
·Detection of more CpGs with fewer sequence reads.
·Uniform dinucleotide distribution.
·High-efficiency library preparation, with larger library insert sizes.
·Conversion module also available separately.