There are various methods for DNA methylation detection, but most of them cannot be separated from bisulfite conversion, which is to convert unmethylated C to U in the sequence. On the other hand, single-molecule real-time sequencing (SMRT) does not require bisulfite treatment but uses DNA polymerase to synthesize and collect fluorescent signals simultaneously. Lifeasible's high throughput method, SMRT, allows you to determine the nucleic acid sequence and the methylation status.

SMRT technology uses fluorescently labeled nucleotides to sequence as they are synthesized, reflecting the polymerization rate of each dNTP in the presence of DNA polymerase by two kinetic parameters, PW and IPD. When DNA polymerase encounters a modifying methyl group of the template strand, such as m6A, m5C, or 5hmc, the rate of dNTP polymerization changes, and PW and PID change accordingly.

• We can identify the type and location of specific DNA template base modifications using these two kinetic parameters, allowing us to analyze DNA methylation profiles at the genome-wide level, with direct and simultaneous detection of m6A, m5C, and m4C.
• Using SMRT, we could determine the methylation profiles of the wild-type and a particular methyltransferase mutant strain of interest, which allowed the discovery of many new features of DNA methyltransferases, such as new recognition sites.

Advantages

• Epigenetics can be analyzed in every run without sulfite treatment.
• Extremely high sequence and methylation accuracy.
• Access to the complete genome and exploration of difficult regions of the genome (e.g., repeats and centromeres) that are inaccessible to short-read long sequencing.
• Identification of allele-specific methylation.

Single-molecule sequencing technology, which reads nucleotide sequences at the level of individual molecules, can generate longer base read lengths, can sequence RNA directly without reverse transcription, and is extremely fast. The ideal method for detecting base modifications should directly apply to natural DNA and not require pre-processing before sequencing. With the in-depth study of epigenetic changes in plants and the further development of high-throughput sequencing technologies, the techniques for detecting DNA methylation continue to improve. Lifeasible offers SMRT to make single-molecule real-time sequencing possible, while providing an opportunity to explore methods for detecting base modifications. Our solution can be applied to regions that are difficult to sequence and can effectively improve the mining and processing of plant DNA methylation data for more biological information. Please feel free to contact us to submit project requirements.

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For research or industrial use.

Related Services

• Restriction Endonuclease-Based DNA Methylation Detection
• Bisulfite-Based DNA Methylation Detection
• Immunoprecipitation of Methylated DNA
• Single Molecule Nanopore Dependent Methylation Sequencing
• RNA Methylation
• RNA-Directed DNA Methylation
• DNA Hydroxymethylation