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EPIGENETICS

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

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.

HistoneModifications

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:

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.

ChIPSequencing
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.

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