The NEBNext Enzymatic Methyl-seq Kit is a commercially available kit from New England Biolabs designed to identify 5-methylcytosine and 5-hydroxymethylcytosine residues from a DNA sample. The protocol makes use of DNA sheared with sonication and converts methylated cytosines through an enzymatic reaction, rather than through bisulfite conversion. This results in higher sensitivity of methylated residues from as little as 10 ng of DNA. A more thorough description of the kit can be found on the NEB website.
Indexes are available for multiplexing of up to 96 samples. Genomic DNA input is 10 ng - 200 ng.
Core staff are available to assist with experimental design and to answer questions related to next-generation sequencing technology. Arrangements for a meeting to discuss new or existing projects can be made by sending an email to Nathan Bivens, Genomics Core director.
All projects must be initiated with a quote before samples are submitted. Send a quote request to the general core email mugenomicscore@missouri.edu.
The Genomic Calculator may be used to estimate the amount of sequence required for a project.
General Considerations:
- Submit samples only after paperwork has been completed.
- Clearly label tubes. Names must match those given on the project information form.
- Low binding microcentrifuge tubes (0.5 or 1.5 mL) are preferred for storing high quality nucleic acids.
Submission Requirements:
| Input Range | Submission Concentration |
Recommended Buffer |
Special Instructions |
| 10-200 ng | 5 - 150 ng/μl DNA | EB buffer; 10 mM Tris-Cl (pH 8.5) |
An image of 100 ng of genomic DNA following agarose gel electrophoresis and staining will be provided by the researcher to demonstrate gDNA integrity. |
| Sequence Depth/Coverage* | Read Type | Index Type | Read Length | ||
| 15x coverage per sample | Paired end | unique dual-index | 150 bases | ||
* minimum coverage recommended by MUGTC; increased coverage may be desired for some studies