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info@allelelifesciences.comMethylation analysis is the study of the chromosomal patterns of DNA or histone modification by methyl groups. This type of modification affects CG dinucleotides and involves the covalent addition of a methyl group (CH3). Lysine residues in histone tails and the cytosine (C) base in DNA can both be methylated.
- COMET Assay
- Methylation Analysis by PCR
- Methylation Analysis by Sequencing
- Analysis by HPLC by estimation of 5-mC or 5-hmC
- Methylation Analysis by ELISA ( measurement of 5-mC or 5-hmC )
COMET Assay:
DNA strand breakage assay also
known as single cell gel electrophoresis assay is a
sensitive and rapid test for quantifying and
analysing DNA damage in individual cells.
DNA Metylation
Analysis by Sequencing :
One of the most commonly used
techniques for detecting DNA methylation
at specific genes or sequences is
methylation-specific PCR. The methylated regions
are then selectively amplified using primers
designed for methylation and sequenced after the
DNA's cytosine is bisulfite-converted to uracil.
In Whole Genome
Bisulfide Sequencing -
Bisulfite conversion PCR primer design PCR
amplification optimization Library preparation
QC of final libraries (DNA concentration,
analysis of the profile) Sequencing on Illumina
platform
Bisulfide
Modification and Methylation HPLC Analysis :
We offer high-performance liquid chromatography
analysis services that are effective and
precise. Exploring the methylation states of
genomic DNA is possible after bisulfite
treatment. Cytosine that has not been methylated
is transformed to uracil and finally to thymine.
It is possible to measure the concentration of 5
mC (5-methylcytosine) on DNA molecules and PCR
products by sequencing and mapping to a
sufficient read depth.
Methylation Analysis by ELISA
The genomic DNA is denatured and fixed, and the methylated cytosines are detected through sequential incubation steps with: (a) a primary antibody raised against 5 mC; (b) a labeled secondary antibody; (c) colorimetric or fluorometric detection reagents; and (d) measurement in a microplate reader. ELISA is a reliable method that can be used to detect epigenetic modifications of DNA immobilised on plastics.