CD4_C8-Treg 3 P1.ut.AAACCTGTCACGGTTA-1 P1 P1.pre CD4 CD4_C8-Treg 4 P1.ut.AAACCTGTCCGCTGTT 1.2 创建seurat对象 依然使用CreateSeuratObject 函数,此处count 为读取的矩阵文件。 1,CreateSeuratObject中的meta.data参数 CreateSeuratObject函数除了简单的过滤条件外 ,还有一个重要的meta.data参数,可以输入提供的meta信息。 CreateSeuratObject函数的帮助文档中也很明确的提到了该点要求。 发现问题后,只需要将meta文件的cellid列转为rownames即可。 sce4 <- sce0 sce4@meta.data <- sce4@meta.data %>% rownames_to_column("cellid") %>% inner_join(meta
Read10X(data.dir = paste(dataset_loc, ids[1],"filtered_feature_bc_matrix", sep="/")) seurat_obj <- CreateSeuratObject Read10X_h5(file.path(dataset_loc, ids[1], "filtered_feature_bc_matrix.h5"), use.names = T) seurat_obj <- CreateSeuratObject , ids[1], "all.datatable.txt"), sep="\t", header=T, row.names=1) # dim(matrix_data) # matrix_data[1:4,1 :4] seurat_obj <- CreateSeuratObject(counts = matrix_data) 方式四:readMM # Read in `matrix.mtx` counts row names as the gene IDs rownames(counts) <- gene_ids colnames(counts) <- cell_ids seurat_obj <- CreateSeuratObject
pbmc500_assay <- CreateChromatinAssay(pbmc500.counts, fragments = frags.500) pbmc500 <- CreateSeuratObject meta.data=md.500) pbmc1k_assay <- CreateChromatinAssay(pbmc1k.counts, fragments = frags.1k) pbmc1k <- CreateSeuratObject meta.data=md.1k) pbmc5k_assay <- CreateChromatinAssay(pbmc5k.counts, fragments = frags.5k) pbmc5k <- CreateSeuratObject meta.data=md.5k) pbmc10k_assay <- CreateChromatinAssay(pbmc10k.counts, fragments = frags.10k) pbmc10k <- CreateSeuratObject features: 0 ## Genome: ## Annotation present: FALSE ## Motifs present: FALSE ## Fragment files: 4
:4] rownames(counts) <- counts[, 1] counts <- counts[, -1] dim(counts) # 创建Seurat对象 sce <- CreateSeuratObject C1 C2 C3 C4 M1 M2 M3 S1 S4 6394 7621 8561 4780 6464 6432 7627 10443 :4] rownames(counts) <- counts[, 1] counts <- counts[, -1] dim(counts) # 创建Seurat对象 sce <- CreateSeuratObject (counts = counts, min.cells=3, project = gsub(".txt","", pro) ) head(sce@meta.data) 在 counts[1:4,1:4 > gsub(".txt","", pro) [1] "IRI1d_1" CreateSeuratObject 这个函数做了什么?
但目前seurat包已经更新到5.0.1版本,更新后使用起来也花了一些时间Seurat包更新与使用初探 虽然感觉在seurat对象结构上,V4和V5版本区别不大——V5和V4版Seurat对象内部结构对比详细版 ,但是在读取数据的时候,V4和V5的区别还是有点明显的。 如果是单个样品,直接读取进来然后创建seurat对象即可:初试Seurat的V5版本 主要区别在于,V4版本中一般是循环读取样品,使用CreateSeuratObject创建seurat对象,然后使用merge ','',pro), colnames(ct) ,sep = '_') ct=ct[,-1] ct[1:4,1:4] return(ct) /GSE184708/GSE184708_raw_counts_gonad_all_samples_XX_XY_E10_to_E16.mtx.gz" ) mtx[1:4,1:4] dim(mtx) cl
H21 <- CreateSeuratObject(counts = H21, project = "H21", min.cells = 3, min.features = 100) H21 H23 <- CreateSeuratObject(counts = H23, project = "H23", min.cells = 3, min.features = 100) H23 H24 <- CreateSeuratObject = H32, project = "H32", min.cells = 3, min.features = 100) H32 H33 <- CreateSeuratObject(counts = H33 = 3, min.features = 100) H36 H38 <- CreateSeuratObject(counts = H38, project = "H38", min.cells = 3, ob.list[[i]]))/125000)) } MACS <- ob.list[[1]] H21 <- ob.list[[2]] H23 <- ob.list[[3]] H24 <- ob.list[[4]
单细胞专题 | 1.单细胞测序(10×genomics技术)的原理 单细胞专题 | 2.如何开始单细胞RNASeq数据分析 单细胞专题 | 3.单细胞转录组的上游分析-从BCL到FASTQ 单细胞专题 | 4. 使用CreateSeuratObject生成Seurat对象,后续分析都是在该对象上进行操作。 csv_files/S01_datafinal.csv", sep=""), header=T, row.names = 1) Sys.time() dim(raw.data) raw.data[1:4,1 :4] head(colnames(raw.data)) # Load metadata metadata <- read.csv(paste(dir,"Data_input/csv_files/S01 }) # 合并: sce_big <- merge(sceList[[1]], y = c(sceList[[2]],sceList[[3]],sceList[[4]
library("Seurat") scrna_data_ctrl <- Read10X("data/GSE96583/ctrl/") ctrl <- CreateSeuratObject( counts , min.cells = 3, min.features = 200) scrna_data_stim <- Read10X("data/GSE96583/stim/") stim <- CreateSeuratObject sample_list){ filedir = str_c("data/GSE96583/",sample) scrna_data <- Read10X(filedir) Seurat_object <- CreateSeuratObject /data" [4] "./data" "./data/GSE96583" "./data/GSE96583" [7] "./data" ". str_c("data/GSE96583/",sample) # 数据的读取 scrna_data <- Read10X(filedir) # 对象的构建 Seurat_object <- CreateSeuratObject
dgCMatrix" # attr(,"package") # [1] "Matrix" # 构建 Seurat 对象 # 初步过滤一般不需要修改参数,除非数据实在太难看 Seurat_object <- CreateSeuratObject ScRNAdata <- Read10X_h5(filename = "GSM3489182_Donor_01_raw_gene_bc_matrices_h5.h5") Seurat_object <- CreateSeuratObject read.table( "data/GSM2829942/GSM2829942_HE6W_LA.TPM.txt", row.names = 1, header = T) Seurat_object <- CreateSeuratObject CreateSeuratObject( counts, project = "CreateSeuratObject", assay = "RNA", names.field = 1,
<-CreateSeuratObject(counts = C149, project = "C149",min.cells = 3, min.features = 200) C152<-CreateSeuratObject 细胞分型 markers = c('TPPP3','KRT18','CD68','FCGR3B','CD1C','CLEC9A','LILRA4','TPSB2','CD3D','KLRD1','MS4A1 ','IGHG4') 很熟悉吧! pDC,TPSB2是Mast cell,CD3D是T cell,KLRD1是NK cell,MS4A1是B cell,IGHG4是Plasma cell,在doublets中表达CD68和CD3D。 ),NK&T中CD3D-T cell(6、10、30),KLRD1-NK cell(18),B中MS4A1-B cell(26),IGHG4-Plasma cell(24,27),others中CD68&
接下来分别读取 library(Seurat) sce.10x <- Read10X(data.dir = '~/four-PBMC-mtx/SRR7722939/') sce1 <- CreateSeuratObject project = "SRR7722939") sce.10x <- Read10X(data.dir = '~/four-PBMC-mtx/SRR7722940/') sce2 <- CreateSeuratObject <- CreateSeuratObject(raw.data = sce.10x, min.cells = 60, SRR7722940" "SRR7722941" "SRR7722942" library(Seurat) sceList = lapply(folders,function(folder){ CreateSeuratObject <- NormalizeData(sce4) sce4 <- ScaleData(sce4, display.progress = F) 找共有的高变异基因 => FindVariableGenes
示例代码是: rm(list=ls()) options(stringsAsFactors = F) library(Seurat) sce1 <- CreateSeuratObject(Read10X AAACGGGGTCCAACTA" [2] "4602STDY7018923___AAAGATGAGGAGTACC" [3] "4602STDY7018923___AAAGATGGTGAACCTT" [4] AACCATGCAGTCGTGC" > head(rl) [1] "RP11-34P13.3_ENSG00000243485" "FAM138A_ENSG00000237613" [3] "OR4F5 也就是说 readMM 函数即可,然后配合CreateSeuratObject来构建对象! 降维聚类分群和生物学注释都走起! 单细胞实战(三) Cell Ranger使用初探 单细胞实战(四) Cell Ranger流程概览 单细胞实战(五) 理解cellranger count的结果 更新版本见:cellranger更新到4啦
(counts = pfc2.data, project = "pfc-demo", min.cells = 3, min.features = 200) pfc3 <- CreateSeuratObject (counts = pfc3.data, project = "pfc-demo", min.cells = 3, min.features = 200) pfc5 <- CreateSeuratObject (counts = pfc5.data, project = "pfc-demo", min.cells = 3, min.features = 200) pfc7 <- CreateSeuratObject ~ 3 .不死心的话,我们不使用SCTtransform,也不去除批次效应,只使用seurat标准流程试试 3#3 标准流程----- head(subset_data@meta.data) All=CreateSeuratObject 4 结论: 虽然不去批次效应也能拿到很好的结果,个人还是建议使用harmony,你觉得呢? 下面来源GPT: 尽管在某些情况下即使不去除批次效应也能得到看似合理的结果,但这可能是偶然的,并且存在风险。
,tsv/txt,h5ad格式10x格式的读取展开代码语言:TXTAI代码解释library(Seurat)ct=Read10X(data.dir="GSE145154_RAW/")seu.obj<-CreateSeuratObject install.packages("hdf5r")ct<-Read10X_h5("GSE200874_RAW/GSM6045826_wt_filtered_gene_bc_matrices_h5_2.h5")seu.obj<-CreateSeuratObject row.names=1#是将第一列设置为行名的意思ct<-read.csv("GSE130148_raw_counts.csv.gz",row.names=1)class(ct)seu.obj<-CreateSeuratObject
tmp = Read10X(file.path(dir,pro )) if(length(tmp)==2){ ct = tmp[[1]] }else{ct = tmp} sce =CreateSeuratObject tmp = Read10X_h5(file.path(dir,pro )) if(length(tmp)==2){ ct = tmp[[1]] }else{ct = tmp} sce =CreateSeuratObject :4] sce=CreateSeuratObject( ct , project = gsub('_gex_raw_counts.rds',' colnames(ct) ,sep = '_') ct=ct[,-1] sce =CreateSeuratObject(counts = ct , colnames(ct) ,sep = '_') ct=ct[,-1] sce =CreateSeuratObject(counts = ct ,
读取h5格式的文件(使用Read10X_h5函数读取h5格式的单细胞数据文件) seurat_data <- Read10X_h5(file = h5_file) # 创建Seurat对象(使用CreateSeuratObject /data/GSE130148/GSE130148_raw_counts.csv.gz"), row.names = 1) # 使用CreateSeuratObject()函数创建Seurat对象,并在此处指定项目名称 seurat_obj <- CreateSeuratObject(counts = seurat_data, min.features /data/GSE130xxx/xxxx.txt.gz"), row.names = 1, header = TRUE, sep = "\t") # 使用CreateSeuratObject()函数创建 Seurat对象,并在此处指定项目名称 seurat_obj <- CreateSeuratObject(counts = seurat_data,
These BAM files were converted back to FASTQ files, then run through Cell Ranger[4] to obtain the count ctrl_raw_feature_bc_matrix") # Turn count matrix into a Seurat object (output is a Seurat object) ctrl <- CreateSeuratObject (counts = ctrl_counts, min.features = 100) 这里我们使用CreateSeuratObject 将矩阵转换为 the tximeta[10] package, while kallisto-bustools output can be read using the BUSpaRse[11] package. 4- term=SRP102802 [4] Cell Ranger: https://support.10xgenomics.com/single-cell-gene-expression/software/
tmp = Read10X(file.path(dir,pro )) if(length(tmp)==2){ ct = tmp[[1]] }else{ct = tmp} sce =CreateSeuratObject sce.all_int.rds') GSE152938$study = 'GSE152938' table(GSE152938$orig.ident) sceList = list( GSE131685 = CreateSeuratObject ( counts = GSE131685@assays$RNA$counts ), GSE152938 = CreateSeuratObject( counts = GSE152938
hackmyvm.eu/1.在官网搜索你想要的镜像,然后下载2.下载好后解压得到.ova的文件,右击选择VMware或者Oracle VirtualBox进行打开3.在弹出的框中,选择存放的位置,然后点击导入4. /n3gr4后面还跟一个php页面。也是得要扫出来的。m414nj3.php然后就是爆破参数。这里ffuf或者抓包都可以,我就选我熟悉的用了。文件包含漏洞,直接弹shell了。 friendster那就可以登录p4l4nc4这个用户了。用私钥登录就好了。登录上去之后直接跑脚本就好了。可以从/etc/passwd提权。那就直接把密码删了就完了。nano改一下就好了。
|-- [298K] features.tsv.gz `-- [ 76M] matrix.mtx.gz 3 directories, 9 files 如果我们按照之前Seurat的V4版本读取 library(data.table) sceList = lapply(samples,function(pro){ # pro=samples[1] print(pro) sce=CreateSeuratObject head(sce.all@meta.data, 10) table(sce.all@meta.data$orig.ident) 上面的代码我写了有好多年了,一直没有更新或者改进,我们依赖于这个V4的版本的 我们这个时候有一个很简单的方法就可以避免分开读取后的merge ,如下所示: tmp = list.dirs('GSE162616_RAW/outputs/')[-1] tmp ct = Read10X(tmp) sce.all=CreateSeuratObject " [3] "GSE162616_RAW/outputs/HCC3" 统一读取成为了一个稀疏矩阵 如果是对函数或者Seurat对象结构不清晰,就会产生如下所示错误的读取方式: > sce.all=CreateSeuratObject