##----设置,包括=假---------------------------------------------------knitr::opts_chunk$set(塌陷=真,注释=“#>”,缓存=FALSE,图宽度=4,图高度=5,图显示=“保持”,global.par=错误)##----epidata,消息=FALSE---------------------------------------------------库(GenomicRanges)图书馆(总结实验)图库(封面)数据(“stackepi”)斯塔克皮##----外挂1----------------------------------------------------------------plotEpistack(打印epistack)(斯塔克皮,assay=“DNAme”,metric_col=“exp”,ylim=c(0,1),zlim=c(0,1),x_labels=c(“-2.5kb”,“TSS”,“+2.5kb”),titles=“DNA甲基化”,图例=“%mCpG”,metric_title=“表达式”,metric_label=“log10(TPM+1)”,metric_transfunc=函数(x)log10(x+1))##----装箱-------------------------------------------------------------------堆栈pi<-addBins(堆栈pi,nbins=5)plotEpistack(打印epistack)(斯塔克皮,assay=“DNAme”,metric_col=“exp”,ylim=c(0,1),zlim=c(0,1),x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),titles=“DNA甲基化”,图例=“%mCpG”,metric_title=“表达式”,metric_label=“log10(TPM+1)”,metric_transfunc=函数(x)log10(x+1))##----颜色------------------------------------------------------------------plotEpistack(打印epistack)(斯塔克皮,assay=“DNAme”,metric_col=“exp”,ylim=c(0,1),zlim=c(0,1),x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),titles=“DNA甲基化”,图例=“%mCpG”,metric_title=“表达式”,metric_label=“log10(TPM+1)”,metric_transfunc=函数(x)log10(x+1),tints=“道奇蓝”,bin_palette=彩虹)##----par,崩溃=真-------------------------------------------------------plotEpistack(打印epistack)(斯塔克皮,assay=“DNAme”,metric_col=“exp”,ylim=c(0,1),zlim=c(0,1),x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),titles=“DNA甲基化”,图例=“%mCpG”,metric_title=“表达式”,metric_label=“log10(TPM+1)”,metric_transfunc=函数(x)log10(x+1),cex=0.4,cex.main=0.6)##----plotAvgerageProfile,图small=TRUE------------------------------------绘图平均配置文件(斯塔克皮,ylim=c(0,1),化验=“DNAme”,x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),)##----plotStackProfile,fig.small=真---------------------------------------打印堆栈配置文件(斯塔克皮,化验=“DNAme”,x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),调色板=hcl.colors,zlim=c(0,1))##----自定义面板-------------------------------------------------------------布局(矩阵(1:3,ncol=1),高度=c(1.5,3,0.5))旧帕尔<-par(mar=c(2.5、4、0.6、0.6))绘图平均配置文件(stackepi,测定=“DNAme”,x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),ylim=c(0,1),)打印堆栈配置文件(stackepi,测定=“DNAme”,x_labels=c(“-2.5kb”,“TSS”,“+2.5kb“),zlim=c(0,1),调色板=hcl.colors)打印堆栈配置文件图例(zlim=c(0,1),调色板=hcl.colors)par(旧par)布局(1)##----示例路径--------------------------------------------------------------路径读取数<-c(rep1=“https://raw.githubusercontent.com/Bioconductor/CSAMA2016/master/lab-5-chipseq/EpigeneticsCSAMA/inst/bedfiles/H3K27ac_rep1_filtered_ucsc_chr6.bed",rep2=“https://raw.githubusercontent.com/Bioconductor/CSAMA2016/master/lab-5-chipseq/EpigeneticsCSAMA/inst/bedfiles/H3K27ac_rep2_filtered_ucsc_chr6.bed",输入=“https://raw.githubusercontent.com/Bioconductor/CSAMA2016/master/lab-5-chipseq/EpigeneticsCSAMA/inst/bedfiles/ES_input_filtered_ucsc_chr6.bed")路径峰值<-c(峰值1=“https://raw.githubusercontent.com/Bioconductor/CSAMA2016/master/lab-5-chipseq/EpigeneticsCSAMA/inst/bedfiles/Rep1_peaks_ucsc_chr6.bed",峰值2=“https://raw.githubusercontent.com/Bioconductor/CSAMA2016/master/lab-5-chipseq/EpigeneticsCSAMA/inst/bedfiles/Rep2_peaks_ucsc_chr6.bed")##----峰值加载,消息=FALSE----------------------------------------------库(rtracklayer)峰值<-重叠(路径峰值,导入)##----峰值合并,消息=假------------------------------------------------merged_peaks<-基因组范围::联合(峰值[1],峰值[2])scores_rep1<-双(长度(合并峰值))scores_rep1[findOverlaps(峰值[1],合并峰值,select=“first”)]<-峰值[1]$scorescores_rep2<-双(长度(合并峰值))scores_rep2[findOverlaps(峰值[2],合并峰值,select=“first”)]<-峰值[2]$score峰值类型<-ifelse(scores_rep1!=0&scores_rep2!=0,“两者”,ifelse(scores_rep1!=0,“仅Rep1”,“仅Rep2”))mcols(合并峰值)<-数据帧(scores_rep1,scores_rep2,peak_type)合并峰值$mean_scores<-apply((mcols(合并峰值)[,c(“scores_rep1”,“scores.rep2”)]),1,mean)merged_peaks<-merged_peaks[顺序(merged_peaks$mean_scores,递减=真),]rm(scores_rep1,scores_rep2,peak_type)合并峰值##----读取_加载-------------------------------------------------------------读取<-lapply(path_reads,import)##----coverage_matrices,消息=FALSE-----------------------------------------图书馆(强化热图)覆盖_面积<-重叠(读取,函数(x){规格化ToMatrix(x、,调整大小(merged_peaks,width=1,fix=“center”),延伸=5000,宽度=250,mean_mode=“覆盖范围”)})xlabs<-c(“-5kb”,“峰值中心”,“+5kb”)##----准备就绪------------------------------------------------------------merged_peaks<-总结实验(rowRanges=合并峰值,分析=覆盖范围)##----setup2,include=假--------------------------------------------------knitr::opts_chunk$set(图宽度=6,图高度=5)##----峰值点----------------------------------------------------------------plotEpistack(打印epistack)(合并峰值,分析=c(“rep1”、“rep2”、“input”),tints=c(“dodgerblue”,“firebrick1”,“grey”),titles=c(“Rep1”、“Rep2”、“Input”),x_labels=xlabs,zlim=c(0,4),ylim=c(0,4),metric_col=“mean_scores”,metric_title=“峰值分数”,metric_label=“分数”)##----峰值_点_bin------------------------------------------------------------rowRanges(merged_peaks)$bin<-rowRangesplotEpistack(打印epistack)(merged_峰值,分析=c(“rep1”、“rep2”、“input”),tints=c(“dodgerblue”,“firebrick1”,“grey”),title=c(“Rep1”、“Rep2”、“输入”),x_labels=xlabs,zlim=c(0,4),ylim=c(0,4),metric_col=“mean_scores”,metric_title=“峰值分数”,metrac_label=“score”,bin_palette=colorRampPalette(c(“深兰色1”,“道奇蓝”,“firebrick1”)),npix_height=300)##----峰值_点_平方英寸-----------------------------------------------------------合并峰值<-合并峰值[顺序(rowRanges(merged_peaks)$bin,rowRange(merged_peaks,递减=c(FALSE,TRUE),方法=“基数”), ]plotEpistack(打印epistack)(合并峰值,模式=c(“rep1”、“rep2”、“input”),tints=c(“dodgerblue”,“firebrick1”,“grey”),titles=c(“Rep1”、“Rep2”、“Input”),x_labels=xlabs,zlim=c(0,4),ylim=c(0,4),metric_col=“mean_scores”,metric_title=“峰值分数”,metrac_label=“score”,bin_palette=colorRampPalette(c(“深兰色1”,“道奇蓝”,“firebrick1”)),npix_height=300)##----示例2_tss-------------------------------------------------------------负载(system.file(“extdata”,“chr21_test_data.RData”,package=“EnrichedHeatmap”),verbose=真)tss<-启动子(基因,上游=0,下游=1)tss$gene_id<-names(tss)总悬浮固体##----表达式数据----------------------------------------------------------expr<-数据帧(gene_id=名称(rpkm),expr=每公里转数)epidata<-addMetricAndArrangeG范围(tss、expr、,gr_key=“gene_id”,order_key=“gene_id”,order_value=“expr”)数据录入##----添加bin--------------------------------------------------------------epidata<-addBins(epidata,nbins=5)数据录入##----信号提取--------------------------------------------------------methstack<-normalizeToMatrix(meth,epidata,value_column=“meth”,extend=5000,w=250,mean_mode=“绝对”)h3k4me3堆栈<-normalizeToMatrix(H3K4me3,epidata,value_column=“覆盖范围”,extend=5000,w=250,mean_mode=“覆盖范围”)epidata<-总结实验(rowRanges=epidata,分析=列表(DNAme=methstack,H3K4me3=H3K4me3 stack))##----示例2_绘图--------------------------------------------------------plotEpistack(打印epistack)(epidata、,tints=c(“减淡蓝”,“橙色”),zlim=列表(c(0,1),c(0,25)),ylim=列表(c0,1,c0,50)),x_labels=c(“-5kb”,“TSS”,“+5kb”),图例=c(“%mCpG”,“覆盖范围”),metric_col=“expr”,metric_title=“基因表达”,metric_label=“log10(RPKM+1)”,metric_transfunc=函数(x)log10(x+1),npix_height=300)##----会话信息--------------------------------------------------------------sessionInfo()