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5mC & 5mhC Sequencing Service


Overview

Epigenetics pertains to hereditary shifts in the expression of genes that are not induced by DNA sequence modifications. Gene expression regulation can happen by post-translation alteration of the amino acids in histone proteins, by DNA methylation, and by microRNAs at the post-transcriptional stage (miRNAs). In order to differentiate invading phage DNA, bacteria utilize an absence of methylation, initiating cleavage by host restriction enzymes. DNA methylation is frequently assumed to suppress gene expression in eukaryotes, although its involvement has lately been revealed to differ considerably by species, the background of sequence and kinds of cell. It has been involved in many cellular and epigenetic mechanisms, such as embryonic development, chromatin structure alteration, inhibition of the X-chromosome, genomic imprinting, preservation of chromosome stability and carcinogenesis. One of DNA methylation's most interesting characteristics is that it can modify in reaction to developmental or environmental indications.

DNA methylation pertains to an addition to a nucleotide of a methyl group. 5-methylcytosine (5mC) is the most familiar type of methylated nucleotide. 5mC can be oxidized to 5-hydroxymethylcytosine (5hmC), which can be identified in almost all mammalian tissues and cells. 5hmC is now regarded as an epigenetic alteration, in relation to being one of the medium states of DNA demethylation.

Our Advantages
  • Evaluate the effect of longer reads, particularly for novel genome de novo assemblies.
  • Assess methyltransferases from DNA.
  • Dependable and precise as more conventional methods.
  • Maximum accuracy of consensus, low sequencing-context bias
  • Initiatives and pipelines for novel bioinformatics evaluation
5mC & 5mhC Sequencing Service
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Introduction to Our Microbial 5mC/5hmC Sequencing Platform

5mC & 5mhC Sequencing Service

Single Molecular Real-Time (SMRT) sequencing uses a customized flow cell with many thousands of transparent bottoms of specific picoliter wells, zero-mode waveguides (ZMW). The polymerase is installed at the bottom of the well and facilitates the progression of the DNA strand through the ZMW. As a result, the platform can concentrate on a single molecular element.SMRT sequencing enables fluorescently tagged nucleotides that are synthesized along with individual DNA layout molecules to be imaged in real-time. When the framework and polymerase disentangle, the sequencing reaction terminates. The PacBio instrument's average read length is roughly 2 kb and some reads may exceed 20 kb. For de novo assemblies of novel genomes that can stretch many more repeats and bases, longer reads are particularly valuable.

5mC & 5mhC Sequencing Service

Single-Molecule, Real-Time (SMRT) Sequencing intercepts DNA shifts by assessing variability in DNA base establishment polymerase kinetics during sequencing, eradicating the requirement for arduous procedures for chemical conversion. With SMRT Sequencing, you have the capacity to:

  • At coverage stages suggested for assembly, identify genome-wide m6A and m4C R-M system themes
  • Evaluate the methylation condition of m6A and m4C at all positions of genomics with marginally greater coverage
  • Acquire complete genomes with epigenetic alteration annotations
  • Expose phase differences of R-M genes that control gene batteries associated in pathogenesis, host adaptation and resistance to antibiotics.

Microbial 5mC/5hmC Sequencing Service Workflow

5mC & 5mhC Sequencing Service

Bioinformatics Analysis

Our bioinformatics analysis includes five parts: Taxonomy Annotation, Gene Prediction, KEGG, eggNOG, and CAZy analyses. Our bioinformatics analysis services are flexible to your specific projects.

ANALYSIS CONTENT DETAILS
Gene Prediction Gene localization along a genome
Taxonomy Annotation In a reference dataset, verify the taxonomy of a sequence
CAZy Assessment of Carbohydrate-Active Enzyme genomic, structural and biochemical data
eggNOG Exhibit relationships in orthology among genes
KEGG Analyzing high-level biological system processes and utilities

Sample Requirements

  • 5mC & 5mhC Sequencing Service

    1. A minimum of 10 μg HMW DNA (samples) is required for long-read sequencing using SMRT.

Sampling kits: We provide a range of microbial sampling kits for clients, including MicroCollect™ oral sample microbial collection products and MicroCollect™ stool sample collection products.

References

  1. Schatz MC. Nanopore sequencing meets epigenetics. Nature methods. 2017, 14(4).
  2. Roberts RJ, Carneiro MO, Schatz MC. The advantages of SMRT sequencing. Genome biology. 2013, 14(7):1-4.
* For Research Use Only. Not for use in diagnostic procedures or other clinical purposes.



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