Phylogenetic and genetic variation analyses are essential methods for understanding and studying species, and obtaining high-quality genome sequence data is an essential prerequisite for such analyses. Nanopore sequencing is a new generation of single-molecule sequencing technology that offers advantages over SMRT in terms of read length (up to 2 Mb for a single read), speed, and real-time monitoring of sequencing data. Lifeasible's single-molecule nanopore sequencing technology enables you to detect endogenous CpG methylation and exogenous labeling of chromatin-accessible sites.

When a single-stranded DNA molecule passes through a nanopore, the different bases have different shapes and sizes and react specifically with the cyclodextrin molecules in the pore, causing a change in resistance. There is a constant voltage on both sides of the nanopore, so changes in the current in the nanopore can be detected, thus reflecting the base arrangement of the DNA molecules passing through the nanopore. Since the resistance changes caused by nucleotides before and after modifications such as methylation are different, we can use this sequencing platform to determine the methylation modification of bases by identifying the electrical signals.

Application case

5mC, the most abundant methylation modification in DNA, is present in most plants and is one of the most representative epigenetic modifications. We provide a methylation sequencing protocol based on single-molecule nanopore-dependent technology that can distinguish between 5mC and normal cytosine in DNA, thus enabling single-base localization analysis of 5mC. In addition, it can be used for localization analysis of 5hmC in DNA molecules.

Advantages

• Signal amplification by PCR is not required during sequencing, thus avoiding base mismatches introduced during the PCR reaction.
• No enzyme catalysis is involved in the entire reaction, so theoretically, as long as the nucleic acid extraction step can obtain a sequence of sufficient length, the sequencing step can detect it.
• The unique sequencing principle of using electrical signals makes it possible to detect base modifications of the four C.

To compensate for the lack of precision, Lifeasible offers a combined strategy of multi-platform sequencing, which allows us to ensure the reliability of genome sequencing results while bringing you methylation information that cannot be found by a single sequencing tool. This brings a new direction for genome sequencing research and provides a possible biological pathway for researchers to study the regulation of plant methylation further and improve crops. Please feel free to contact us to submit your requirements and get our updated proposal.

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For research use only, not intended for any clinical use.

Related Services

• Restriction Endonuclease-Based DNA Methylation Detection
• Bisulfite-Based DNA Methylation Detection
• Immunoprecipitation of Methylated DNA
• SMRT Sequencing for Direct Detection of DNA Methylation
• RNA Methylation
• RNA-Directed DNA Methylation
• DNA Hydroxymethylation