The term "Epigenetics" describe all covalent DNA modifications which impact gene expression without affecting genetic sequence. Epigenetics helps to understand how the environment might impact our gene expression and allow to dissect some of the major biological processes such as aging, cancer and the development of deseases. The progress of epigenetic products development over the years has improvede the detection of epigenetic modifications making easier to gain a significant insight of samples and biological processes
Epigenetics Research look after different "events":
DNA methylation is the addition of a methyl group to DNA, specifically cytosine, by an enzyme known as DNA methyltransferase (DNMT), thereby affecting gene expression. This epigenetic mark typically reduces the expression of genes.
DNA Methylation may be evaluated using different approaches including:
1) Global DNA Methylation
The easiest way to begin research in epigenetics is by investigating global DNA methylation. Measuring total 5-methylcytosine (5-mC) levels may reveal possible epigenetic factors at play and offer invaluable insight into your samples. It has been implicated as a biomarker in numerous human cancers and could help assess neurodegenerative diseases such asAlzheimer's and Parkinson's.
Directly quantifying total 5-mC for an accurate measurement of global DNA methylation can be accomplished in under 4 hours using the MethylFlashMethylated (5-mC) DNA Quantification Kit, a simple ELISA-like method.
2) Methylation-specific PCR (MSP)
The gene-specific, base-resolution DNA methylationanalysisassesses the amount of DNA methylation of a specific gene at base resolution, compared to global DNA methylation, whichprovides an overallview of the entiremethylation status of the genome. The BisulFlash DNA Modification Kit is the ideal kit for quickbisulfite treatment of DNA gearedtowardsmethylation-specific PCR analysis. MSP begins with bisulfiteconversionwhereinunmethylatedcytosineisconverted to uracil. In subsequent PCR amplification, uracilisrecognizedasthymine, leaving the methylatedcytosines (5-mC) unaffected. Thisallows the amount of methylation to be measured.
3) DNA BisulfiteSequencing (Methyl-Seq)
Common DNA methylation-basedapplications include wholegenome (WGBS), reducedrepresentation (RRBS), and targetedbisulfite-seq. Samples can be sequenced on an Illumina instrument for precise identification of methylatedgenes of the sample. The EpiNext High-SensitivityBisulfite-Seq Kit (Illumina) isideal for streamlinedbisulfitesequencing NGS librarypreparation from low input DNA. It’s an all-in-one kit so itincludeseverythingneeded for bisulfiteconversion and librarypreparation.
Post-translational modifications (PTMs) which occur to histones are known to play a crucial role in transcription repression or activation. Such histone modifications include histone methylation, histone acetylation, and histone phosphorylation.
Global H3 or H4 Modification ELISA constitutes a useful tool to quantify H3 or H4 specific modifications
It is possible to investigate the occurrence of 21 different histone H3 modifications or 10 different histone H4 modifications on one microplate using the EpiQuikHistone H3 Modification Multiplex Assay Kit or the EpiQuikHistone H4Modification Multiplex Assay Kit.
Specific single histone methylation or histone acetylation modification ca be measure using specific kit:
- EpiQuik Global Tri-MethylHistone H3K27 Quantification Kit (Colorimetric)
- EpiQuik Global AcetylHistone H3K9 Quantification Kit (Colorimetric)
- EpiQuik Global Histone H3K27 MethylationAssay Kit
- EpiQuik Global Histone H3AcetylationAssay Kit
Chromatin Immunoprecipitation- ChIP
Chromatin Immunoprecipitation, or ChIP allows to investigating protein-DNA binding interactions. It can be used to identify histone modifications at the gene-specific level. ChIP cosnecutive applications include ChIP-sequencing, ChIP-PCR, and ChIP-on-Chip (microarrays). The ChromaFlash High-Sensitivity ChIP Kit allows the immunoprecipitation of chromatin (ChIP) from small amounts of mammalian cells or tissues.
This method combines chromatin immunoprecipitation (ChIP) with massively parallel sequencing technology to precisely analyze protein interactions with DNA. ChIP-Seq is used for pinpointing the exact gene binding location of the protein of interest.
The EpiNextChIP-Seq High-Sensitivity Kit (Illumina) is ideal to perform ChIP and librarypreparation with a single kit.