E7120S, NEBNext Enzymatic Methyl-seq Kit - 24 assays

The NEBNext Enzymatic Methyl-seq Kit provides a high-performance enzyme-based alternative to bisulfite conversion for methylome analysis using Illumina^® sequencing. 

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.

NEBNext UltraShear(NEB #M7634) has been optimized for enzymatic fragmentation of DNA compatible with EM-seq workflows, and protocols are available in the NEBNext UltraShear product manual.

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

For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Kit (NEB #E3350) is now also available.

NEBNext UltraShear can be used for enzymatic DNA fragmentation in the EM-seq workflow.