Analysis of the methylome of human embryonic stem cells employing methylated DNA immunoprecipitation coupled to next-generation sequencing.
نویسندگان
چکیده
The analysis of DNA-methylation on a genome-wide scale by next-generation sequencing techniques is an invaluable tool towards the understanding of the epigenetic basis of cellular differentiation. Methylated DNA immunoprecipitation (MeDIP) is an immunocapturing method using an antibody targeting 5-methylcytidine (5 mC) and thereby enriching methylated DNA. MeDIP combined with next-generation sequencing (MeDIP-seq) provides a powerful tool for the analysis of genome-wide DNA-methylation profiles. Here, we describe a protocol for the preparation of MeDIP samples suitable for next-generation sequencing on a Genome Analyser (Illumina).
منابع مشابه
O-11: N-a-acetyltransferase 10 Protein Regulates DNA Methylation and Embryonic Development
Background Genomic imprinting is a heritable and developmentally essential phenomenon by which gene expression occurs in an allele-specific manner1. While the imprinted alleles are primarily silenced by DNA methylation, it remains largely unknown how methylation is targeted to imprinting control region (ICR), also called differentially methylated region (DMR), and maintained. Here we show that ...
متن کاملSingle-cell methylome landscapes of mouse embryonic stem cells and early embryos analyzed using reduced representation bisulfite sequencing.
DNA methylation is crucial for a wide variety of biological processes, yet no technique suitable for the methylome analysis of DNA methylation at single-cell resolution is available. Here, we describe a methylome analysis technique that enables single-cell and single-base resolution DNA methylation analysis based on reduced representation bisulfite sequencing (scRRBS). The technique is highly s...
متن کاملChromatin immunoprecipitation-based analysis of gene regulatory networks operative in human embryonic stem cells.
Chromatin immunoprecipitation (ChIP) followed by microarray-based (ChIP-Chip) or next-generation sequencing-based (ChIP-Seq) analysis has been established as a powerful and widely used method to investigate DNA-protein interactions relative to a genomic location in vivo. Here, we present a ChIP-Chip protocol, which utilizes an alternative, easier amplification protocol and when using high-quali...
متن کاملComputational analysis of genome-wide DNA methylation during the differentiation of human embryonic stem cells along the endodermal lineage.
The generation of genome-wide data derived from methylated DNA immunoprecipitation followed by sequencing (MeDIP-seq) has become a major tool for epigenetic studies in health and disease. The computational analysis of such data, however, still falls short on accuracy, sensitivity, and speed. We propose a time-efficient statistical method that is able to cope with the inherent complexity of MeDI...
متن کاملI-37: Establishing High Resolution Genomic Profiles of Single Cells Using Microarray and Next-Generation Sequencing Technologies
The nature and pace of genome mutation is largely unknown. Standard methods to investigate DNA-mutation rely on arraying or sequencing DNA from a population of cells, hence the genetic composition of individual cells is lost and de novo mutation in cell(s) is concealed within the bulk signal. We developed methods based on (SNP-) arraying and next-generation sequencing of single-cell whole-genom...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Methods in molecular biology
دوره 873 شماره
صفحات -
تاریخ انتشار 2012