New England Biolabs (NEB®) at present introduced the launch of the NEBNext Enzymatic 5hmC-seq Equipment (E5hmC-seq™), a novel enzyme-based technique for the particular detection of 5hmC websites. The light, enzyme-based method permits 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 vital position in gene regulation and different organic processes.
“Thus far, the examine of 5hmC has been hampered by the dearth of exact detection strategies,” mentioned Fiona Stewart, Affiliate Director, NEBNext Portfolio Administration. “Whereas NEBNext Enzymatic Methyl-seq (EM-seq™), our gold normal for methylation detection, detects each 5mC and 5hmC, it doesn’t distinguish between them. Moreover, bisulfite-based strategies undergo from lowered knowledge high quality as a result of 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 particular detection of 5hmC websites utilizing a two-step enzymatic conversion workflow,” mentioned Stewart. “The enzymatic technique minimizes DNA harm 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 consists of 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 be out there, for functions past library prep.
For extra info, go to www.NEBNext.com.