​New England Biolabs Launches EM-seq v2, an Updated Kit for More Sensitive Detection of 5mC and 5hmC

IPSWICH, MA, JANUARY 6, 2025—New England Biolabs (NEB) announces the launch of EM-seq v2, a high-performance enzyme-based alternative to bisulfite sequencing for the identification of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). The enzyme-based conversion workflows for EM-seq and E5hmC-seq avoid the pitfalls of whole genome bisulfite treatment, including DNA damage that results in fragmentation, loss, and bias.

The newly updated kit delivers improved performance for wider input range (100 pg to 200 ng), and a faster, more streamlined workflow as compared to the original EM-seq kit.

“EM-seq v2 will be particularly valuable for researchers working in clinical research and epigenetics who often face challenges sequencing samples less than 10 ng that are below the input range of bisulfite sequencing and our original EM-seq kit,” says Andrew Barry, associate director of product portfolio management for NEB’s sequencing product line. “The kit’s high-quality data from low input samples will enable researchers to get improved data for the same amount of sequencing or similar data with less sequencing.”

Key improvements in EM-seq v2 include:

• Substantially lower DNA input: The minimum input requirement has been reduced from 10 ng to 100 picograms, making EM-seq v2 suitable for applications with limited sample availability.
• Streamlined workflow: EM-seq v2 features a faster incubation step and eliminates a cleanup step, resulting in a time savings of approximately 30 to 45 minutes.
• Improved performance: The kit delivers enhanced data quality, with greater mapping efficiency, more even GC coverage, and detection of more CpGs with fewer sequence reads.

“As an ISO 17025 accredited laboratory, we need reliable and reproducible kits and protocols,” says Dr. Marta Gut, head of the sequencing unit at Centro Nacional de Análisis Genómico in Barcelona, Spain. “After thorough benchmarking of NEBNext Enzymatic Methyl-Seq v2 interrogating 5mC, the standard bisulfite conversion method lost its reference status for our methylation analyses. NEBNext EM-seq v2 and the NEBNext E5hmC-seq kit for detecting 5hmC, specifically, are now our methods of choice to study both modifications.”

EM-seq v2 is compatible with automation for easy scale-up and with 5hmC-seq for specific detection of 5hmC. Indexing primers are now supplied separately, providing greater flexibility for multiplexing samples. For further streamlining, enzymatic fragmentation of DNA compatible with EM-seq and E5hmC-seq workflows can be achieved using NEBNext UltraShear.

The original EM-seq kit is not being discontinued at this time. More details and extensive performance data can be found at www.NEBNext.com.

- This press release was originally published by New England Biolabs.