New England Biolabs (NEB®) as we speak introduced the launch of the NEBNext Enzymatic 5hmC-seq Equipment (E5hmC-seq™), a novel enzyme-based technique for the precise detection of 5hmC websites. The mild, enzyme-based strategy allows excessive yields and high-quality knowledge, with an enter vary of 100 pg to 200 ng.
Whereas the organic significance of 5hmC modification is much less clear than that of 5mC, the abundance of 5hmC varies considerably between tissues, suggesting that it might play a crucial function in gene regulation and different organic processes.
“Up to now, the research of 5hmC has been hampered by the shortage of exact detection strategies,” mentioned Fiona Stewart, Affiliate Director, NEBNext Portfolio Administration. “Whereas NEBNext Enzymatic Methyl-seq (EM-seq™), our gold commonplace for methylation detection, detects each 5mC and 5hmC, it doesn’t distinguish between them. Moreover, bisulfite-based strategies endure from diminished knowledge high quality because of the pattern fragmentation and lack of DNA that outcomes from the damaging bisulfite remedy, thereby limiting their sensible utility.
“To handle these challenges, we developed the NEBNext Enzymatic 5hmC-seq Equipment, which permits for the precise detection of 5hmC websites utilizing a two-step enzymatic conversion workflow,” mentioned Stewart. “The enzymatic technique minimizes DNA injury and permits for the discrimination of 5hmC from unmodified cytosine and 5mC after Illumina® sequencing. Moreover, E5hmC-seq knowledge might be subtracted from EM-seq knowledge, permitting for exact dedication of particular person 5mC and 5hmC websites.”
The equipment contains the reagents required for E5hmC-seq conversion and library preparation suitable with Illumina sequencing; index primers for multiplexing can be found individually. A conversion module can also be obtainable, for functions past library prep.
For extra info, go to www.NEBNext.com.
