Methylome analysis using MeDIP-seq with low DNA concentrations |
| |
Authors: | Taiwo Oluwatosin Wilson Gareth A Morris Tiffany Seisenberger Stefanie Reik Wolf Pearce Daniel Beck Stephan Butcher Lee M |
| |
Affiliation: | University College London (UCL) Cancer Institute, University College London, London, UK. t.taiwo@ucl.ac.uk |
| |
Abstract: | DNA methylation is an epigenetic mark that has a crucial role in many biological processes. To understand the functional consequences of DNA methylation on phenotypic plasticity, a genome-wide analysis should be embraced. This in turn requires a technique that balances accuracy, genome coverage, resolution and cost, yet is low in DNA input in order to minimize the drain on precious samples. Methylated DNA immunoprecipitation-sequencing (MeDIP-seq) fulfils these criteria, combining MeDIP with massively parallel DNA sequencing. Here we report an improved protocol using 100-fold less genomic DNA than that commonly used. We show comparable results for specificity (>97%) and enrichment (>100-fold) over a wide range of DNA concentrations (5,000-50 ng) and demonstrate the utility of the protocol for the generation of methylomes from rare bone marrow cells using 160-300 ng of starting DNA. The protocol described here, i.e., DNA extraction to generation of MeDIP-seq library, can be completed within 3-5 d. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|