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.
For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Conversion Module (NEB #E3365) is now also available.
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.
Features:
For specific detection of 5hmC, the NEBNext Enzymatic E5hmC-seq Conversion Module (NEB #E3365) is now also available.
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
E7125S | -20 |
Control DNA CpG methylated pUC19 | E7122AVIAL | -20 | 1 x 0.024 ml | 0.1 ng/µl | |
Control DNA CpG unmethylated Lambda | E7123AVIAL | -20 | 1 x 0.024 ml | 2 ng/µl | |
Elution Buffer | E7124AVIAL | -20 | 1 x 2.1 ml | ||
TET2 Reaction Buffer | E7126AVIAL | -20 | 1 x 0.3 ml | ||
TET2 Reaction Buffer Supplement | E7127AVIAL | -20 | 3 x 100 µl | 100 X | |
Oxidation Supplement | E7128AVIAL | -20 | 1 x 0.024 ml | ||
Oxidation Enhancer | E7129AVIAL | -20 | 1 x 0.024 ml | ||
TET2 | E7130AVIAL | -20 | 1 x 0.096 ml | ||
Fe (II) Solution | E7131AVIAL | -20 | 1 x 0.024 ml | ||
Stop Reagent | E7132AVIAL | -20 | 1 x 0.024 ml | ||
APOBEC | E7133AVIAL | -20 | 1 x 0.024 ml | ||
APOBEC Reaction Buffer | E7134AVIAL | -20 | 1 x 0.24 ml | ||
BSA | E7135AVIAL | -20 | 1 x 0.024 ml | ||
DTT | E7139AAVIAL | -20 | 1 x 0.5 ml | 100 mM |
E7125L | -20 |
Control DNA CpG methylated pUC19 | E7122AAVIAL | -20 | 1 x 0.096 ml | 0.1 ng/µl | |
Control DNA CpG unmethylated Lambda | E7123AAVIAL | -20 | 1 x 0.096 ml | 2 ng/µl | |
Elution Buffer | E7124AAVIAL | -20 | 1 x 8.6 ml | ||
TET2 Reaction Buffer | E7126AAVIAL | -20 | 1 x 1.2 ml | ||
TET2 Reaction Buffer Supplement | E7127AAVIAL | -20 | 3 x 400 µl | 100 X | |
Oxidation Supplement | E7128AAVIAL | -20 | 1 x 0.096 ml | ||
Oxidation Enhancer | E7129AAVIAL | -20 | 1 x 0.096 ml | ||
TET2 | E7130AAVIAL | -20 | 1 x 0.384 ml | ||
Fe (II) Solution | E7131AAVIAL | -20 | 1 x 0.096 ml | ||
Stop Reagent | E7132AAVIAL | -20 | 1 x 0.096 ml | ||
APOBEC | E7133AAVIAL | -20 | 1 x 0.096 ml | ||
APOBEC Reaction Buffer | E7134AAVIAL | -20 | 1 x 0.96 ml | ||
BSA | E7135AAVIAL | -20 | 1 x 0.096 ml | ||
DTT | E7139AAVIAL | -20 | 1 x 0.5 ml | 100 mM |
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.In the United States and Europe, this product cannot be used in, or incorporated into, any blood-based, sequencing-based screening diagnostic assay (including product or service-based offerings) for the early detection of colorectal, gastric, lung, liver, ovarian, breast, prostate, esophageal, thyroid, uterine, pancreatic, bladder or kidney cancer indications.