NOMe-Seq Kit

NOMe-Seq is a first of its kind kit to simultaneously analyze nucleosome occupancy and CpG methylation status on the same DNA strand. NOMe-Seq (Nucleosome Occupancy and Methylome sequencing) is a published method that was developed by the Peter Jones lab at USC and has been validated for gene-specific analysis using both traditional cloning and Sanger sequencing methods as well as Next-Gen sequencing. The advantage of NOMe-Seq is the ability to study multiple epigenetic modifications on the same DNA molecule. Traditional methods have only been able to look individually at DNA methylation (bisulfite sequencing) or at nucleosome positioning (MNase-seq or FAIRE-seq) without the ability to understand the relationship between the two.

The kit is useful to study the relationship between nucleosome occupancy, transcription factor binding and DNA methylation within the same DNA molecule. It provides a temporal relationship between nucleosome occupancy and DNA methylation. Due to the high level of resolution that can be obtained from GpC distribution, even subtle changes in nucleosome position can be determined. With the use of fixed chromatin, footprinting information regarding the presence of non-nucleosomal DNA binding proteins (transcription factors) can be obtained. This technology can also be used to identify novel imprinted genes. Finally, NOMe-Seq also offers a tool for researchers to use in studying the different states of chromatin and its effect on gene regulation.

Features:

• Formaldehyde fixation of cells
• GpC Methyltransferase treatment
• Includes chromatin preparation reagents, GpC Methyltransferase enzyme, bisulfite conversion reagents and DNA purification reagents
• Recommendations for cloning and sequencing are available

Benefits:

• Preserves protein/DNA interactions enabling the identification of nucleosome and transcription factor binding sites.
• The high density of GpC dinucleotides in the genome allows for high resolution of nucleosome positioning without compromising identification of endogenous CpG methylation
• Entire workflow is included in the kit from sample preparation to DNA purification.
• Validated downstream analysis verified with known gene targets using both Sanger and Next-gen sequencing