Rapid and efficient preparation of DNA, proteins, and chromatin is vital in performing downstream analyses. Our suite of sample preparation kits are suitable for nearly any workflow, whether it be isolating chromatin for ChIP-Seq or ChIP-PCR applications or preparing DNA from a variety of starting materials for Bisulfite-Seq or MS-PCR.
Our sample preparation kits enable you to go directly from extraction to reaction, offering countless possibilities and flexibility in your experimental workflow. Our DNA, protein, and chromatin preparation kits are the starting point for endless research opportunities and downstream applications.
|Next Generation Sequencing|
Next generation sequencing technologies have increased data output volumes and lowered the cost of DNA sequencing beyond what is possible with standard dye-terminator methods. This has lead to a significant advancement of understanding of several epigenetic mechanisms, namely genome-wide DNA methylation and protein-DNA interaction. Epigentek offers many inexpensive solutions for next-generation sequencing including full package kits to conveniently construct high quality DNA libraries ...
DNA methylation is a major epigenetic modification involving the addition of a methyl group to the 5 position of cytosine by DNA methyltransferase to form 5-methylcytosine (5-mC). This epigenetic mark has the power to turn genes on or off and can be inherited through cell division. DNA methylation was the first epigenetic mark to be discovered and it plays an important role in normal human development, aging, tumorigenesis, and other genetic and epigenetic diseases. Similarly, DNA hydroxymethylation, caused by oxidation of 5-mC through the TET family of enzymes, was further discovered to be involved in controlling gene expression and is a key DNA demethylation process as part of the DNA methylation cycle.
Techniques to measure 5-mC have developed over the years and can vary from heat-based to chemical-based methods. Bisulfite conversion is a popular technique used for preparing DNA for methylation analysis on a gene-specific level. This process converts cytosine to uracil while leaving 5-methylcytosine intact to allow methylation analysis at single-nucleotide resolution. Not only is efficient conversion necessary, but recovery must be optimal due to the harsh chemical reaction which often degrades the DNA.
Obtaining an overall view of a sample’s methylation status is a smart way to pre-screen your samples before pursuing downstream applications. Our MethylFlash global DNA methylation pre-screening tools can be used prior to more in depth investigation and costly sequencing. Conventional methods such as mass spectrometry or chromatography, which require expensive equipment or specialized expertise, are not necessary for this ELISA-based method. Also, unlike LUMA or LINE-1, Alu, and LTR-based assays, MethylFlash technology directly quantifies actual global DNA methylation.
Additional methods for investigating DNA methylation include an antibody-based technique known as methylated DNA immunoprecipitation (MeDIP) that is used for studying gene-specific DNA methylation on a genome-wide scale, activity/inhibition assays that measure DNA methyltransferase (DNMT) and other enzymes related to the process of DNA methylation, as well as an amplification technique involving methylation-specific PCR (MS-qPCR) that is used for studying gene or sequence-specific DNA methylation.
RNA methylation is a reversible post-translational modification to RNA that epigenetically impacts numerous biological processes. It occurs in different RNAs including tRNA, rRNA, mRNA, tmRNA, snRNA, snoRNA, miRNA, and viral RNA. Different catalytic strategies are employed for RNA methylation by a variety of RNA-methyltransferases.
|Chromatin & Transcription|
Chromatin structures are regulated by various mechanisms including histone modification and chromatin remodeling, which involve the binding of transcription factors. By using tools such as chromatin immunoprecipitation, it is possible to gain further insight into the dynamic interactions between transcription proteins and components of chromatin, and to ultimately understand their roles in cellular fu...
Histone methylation causes transcription repression or activation, depending on the target sites. Histone methyltransferases (HMTs) control or regulate DNA methylation through chromatin-dependent transcription repression or activation. Measurement of histone methyltrasferase activity and quantification of histone methylation patterns have become pivotal in studying epigenetic regulation of genes, as well as inhibitor discovery.
|Acetylation & Deacetylation|
Histone acetylation and histone deacetylation involve the addition or removal of an acetyl group on lysine residues in the N-terminal tail and on the surface of the nucelosome core of histone proteins. Acetylated and deacetylated histones are considered epigenetic tags within chromatin by relaxing (euchromatin) or tightening (heterochromatin) chromatin structure, subsequently increasing or decreasing gene transcription levels.
|DNA Damage & Repair|
Direct DNA damage can occur due to a number of factors including oxidative stress, cellular exposure to genotoxic agents, and exposure to cytotoxic agents. Oxidative stress has been shown to be the greatest factor in DNA damage, and increased levels have been closely correlated to harmful environmental factors such as ionizing radiation, industrial chemicals, air pollution, cigarette smoking, and cancer chemotherapy. Measurement of DNA damage will help to further understa...
|Gene Editing & Silencing|
Targeted gene knockdown using small interfering RNA (siRNA) or antisense oligonucleotides has been valuable technology in studying gene function. Gene knockdown leads to the reduction of messenger RNA and subsequently decreased protein expression. Alternatively, CRISPR/Cas9-based gene editing is also a versatile method to alter the genome. These gene editing and silencing methods give researchers valuable insight into biological processes and especially diseases.
|Other Histone Modifications|
Various histone modifications, such as histone citrullination and histone phosphorylation, have been shown to epigenetically impact gene expression through different mechanisms. Adjusting epigenetic marks and changing chromatin structure – known as chromatin remodeling – as a result of these histone modifications can influence gene regulation in many tissue types. Studying these histone modifications provides researchers with valuable insight into cellular processes such as apoptosis, cel...