NEBNext® Enzymatic Methyl-seq (EM-seq™) is a new method for identification of 5-mC and 5-hmC.
While bisulfite sequencing has been the gold standard for methylome analysis, this conversion treatment damages DNA, resulting in fragmentation, loss and bias. The NEBNext Enzymatic Methyl-seq Conversion Module provides an enzyme-based alternative to bisulfite conversion.
For use in Illumina® sequencing workflows, we recommend use of the NEBNext Enzymatic Methyl-seq Kit (NEB #E7120) instead of this module. The full kit includes the EM-seq Adaptor, which has been optimized for high performance in the EM-seq workflow, as well as the high-efficiency Ultra II library prep reagents, and these components are not included in the Conversion Module.
The NEBNext Enzymatic Methyl-seq Conversion Module provides a high-performance enzyme-based alternative to bisulfite conversion for methylome analysis.
In the EM-seq workflow, TET2 oxidizes 5-mC and 5-hmC, providing protection from deamination by APOBEC in the next step. In contrast, unmodified cytosines are deaminated to uracils.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|Control DNA CpG methylated pUC19||-20|
|Control DNA CpG unmethylated Lambda||-20|
|TET2 Reaction Buffer||-20|
|TET2 Reaction Buffer Supplement||-20|
|Fe (II) Solution||-20|
|APOBEC Reaction Buffer||-20|
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