Abstract

DNA methylation, one of the most important epigenetic modifications, plays a crucial role in various biological processes. The level of DNA methylation can be measured using whole-genome bisulfite sequencing at single base resolution. However, until now, there is a paucity of publicly available software for carrying out integrated methylation data analysis. In this study, we implemented Methy-Pipe, which not only fulfills the core data analysis requirements (e.g. sequence alignment, differential methylation analysis, etc.) but also provides useful tools for methylation data annotation and visualization. Specifically, it uses Burrow-Wheeler Transform (BWT) algorithm to directly align bisulfite sequencing reads to a reference genome and implements a novel sliding window based approach with statistical methods for the identification of differentially methylated regions (DMRs). The capability of processing data parallelly allows it to outperform a number of other bisulfite alignment software packages. To demonstrate its utility and performance, we applied it to both real and simulated bisulfite sequencing datasets. The results indicate that Methy-Pipe can accurately estimate methylation densities, identify DMRs and provide a variety of utility programs for downstream methylation data analysis. In summary, Methy-Pipe is a useful pipeline that can process whole genome bisulfite sequencing data in an efficient, accurate, and user-friendly manner. Software and test dataset are available at http://sunlab.lihs.cuhk.edu.hk/methy-pipe/.

Highlights

  • DNA methylation is a biochemical process that predominantly involves the addition of a methyl group to cytosine nucleotides by DNA methyltransferases

  • Methylation data analysis is conducted through two consecutive software modules: (i) BSAligner module, a bisulfite sequencing read alignment module for data pre-processing and sequence alignment that is implemented based on 2BWT [17] source code and SOAP2 [18] source code; (ii) BSAnalyzer module, a data analysis module implemented to provide a variety of functionalities to facilitate the downstream methylation data analysis

  • To demonstrate the functionality and usage of Methy-Pipe, we applied it to a whole genome bisulfite sequencing dataset from our previous study [10]

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Summary

Introduction

DNA methylation is a biochemical process that predominantly involves the addition of a methyl group to cytosine nucleotides by DNA methyltransferases. To overcome this limitation and gain genome-wide coverage for CpGs, other bisulfite sequencing protocols such as MethylC-Seq [3] and BS-Seq [2] have been developed. These two protocols mainly differ in their amplification procedures. Some software packages are designed for bisulfite sequencing read alignment only [11,12,13,14], others are for specific downstream analysis [15,16] To fill this gap, we implemented Methy-Pipe, an integrative bioinformatics software package that meets the core methylation data analysis demands and provides a variety of analysis tools to facilitate the downstream analysis in an efficient and integrative manner

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