scholarly journals Comprehensive Evaluation of Differential Methylation Analysis Methods for Bisulfite Sequencing Data

Author(s):  
Yongjun Piao ◽  
Wanxue Xu ◽  
Kwang Ho Park ◽  
Keun Ho Ryu ◽  
Rong Xiang

Background: With advances in next-generation sequencing technologies, the bisulfite conversion of genomic DNA followed by sequencing has become the predominant technique for quantifying genome-wide DNA methylation at single-base resolution. A large number of computational approaches are available in literature for identifying differentially methylated regions in bisulfite sequencing data, and more are being developed continuously. Results: Here, we focused on a comprehensive evaluation of commonly used differential methylation analysis methods and describe the potential strengths and limitations of each method. We found that there are large differences among methods, and no single method consistently ranked first in all benchmarking. Moreover, smoothing seemed not to improve the performance greatly, and a small number of replicates created more difficulties in the computational analysis of BS-seq data than low sequencing depth. Conclusions: Data analysis and interpretation should be performed with great care, especially when the number of replicates or sequencing depth is limited.

2017 ◽  
Author(s):  
Katarzyna Wreczycka ◽  
Alexander Gosdschan ◽  
Dilmurat Yusuf ◽  
Björn Grüening ◽  
Yassen Assenov ◽  
...  

AbstractDNA methylation is one of the main epigenetic modifications in the eukaryotic genome; it has been shown to play a role in cell-type specific regulation of gene expression, and therefore cell-type identity. Bisulfite sequencing is the gold-standard for measuring methylation over the genomes of interest. Here, we review several techniques used for the analysis of high-throughput bisulfite sequencing. We introduce specialized short-read alignment techniques as well as pre/post-alignment quality check methods to ensure data quality. Furthermore, we discuss subsequent analysis steps after alignment. We introduce various differential methylation methods and compare their performance using simulated and real bisulfite sequencing datasets. We also discuss the methods used to segment methylomes in order to pinpoint regulatory regions. We introduce annotation methods that can be used for further classification of regions returned by segmentation and differential methylation methods. Finally, we review software packages that implement strategies to efficiently deal with large bisulfite sequencing datasets locally and we discuss online analysis workflows that do not require any prior programming skills. The analysis strategies described in this review will guide researchers at any level to the best practices of bisulfite sequencing analysis.


2008 ◽  
Vol 36 (5) ◽  
pp. e34-e34 ◽  
Author(s):  
C. Rohde ◽  
Y. Zhang ◽  
T. P. Jurkowski ◽  
H. Stamerjohanns ◽  
R. Reinhardt ◽  
...  

2017 ◽  
Vol 19 (5) ◽  
pp. 737-753 ◽  
Author(s):  
Adib Shafi ◽  
Cristina Mitrea ◽  
Tin Nguyen ◽  
Sorin Draghici

Author(s):  
Xiaoqing Yu ◽  
Shuying Sun

AbstractWe are presenting a comprehensive comparative analysis of five differential methylation (DM) identification methods: methylKit, BSmooth, BiSeq, HMM-DM, and HMM-Fisher, which are developed for bisulfite sequencing (BS) data. We summarize the features of these methods from several analytical aspects and compare their performances using both simulated and real BS datasets. Our comparison results are summarized below. First, parameter settings may largely affect the accuracy of DM identification. Different from default settings, modified parameter settings yield higher sensitivity and/or lower false positive rates. Second, all five methods show more accurate results when identifying simulated DM regions that are long and have small within-group variation, but they have low concordance, probably due to the different approaches they have used for DM identification. Third, HMM-DM and HMM-Fisher yield relatively higher sensitivity and lower false positive rates than others, especially in DM regions with large variation. Finally, we have found that among the three methods that involve methylation estimation (methylKit, BSmooth, and BiSeq), BiSeq can best present raw methylation signals. Therefore, based on these results, we suggest that users select DM identification methods based on the characteristics of their data and the advantages of each method.


2008 ◽  
Vol 2008 (Spring) ◽  
Author(s):  
Christian Rohde ◽  
Yingying Zhang ◽  
Tomasz P. Jurkowski ◽  
Heinrich Stamerjohanns ◽  
Richard Reinhardt* ◽  
...  

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