Comments (3)
Installation
Install proper packages
# 1. Install function libraries ------------------------
if (FALSE) {
"Run these only once"
install.packages(tidyverse)
install.packages(cowplot)
install.packages(Seurat)
install.packages(tictoc)
install.packages("devtools"); # If you don't have it
require("devtools")
devtools::install_github(repo = "vertesy/MarkdownReportsDev")
}
Installation for custom function libraries is not needed.
Installation is only needed for packages. You only need to load these libraries with the source()
command.
There are two ways to load them
source()
directly from the web (recommended)- Download locally and
source()
it from the folder you downloaded to.
You will need these custom function libraries
If the installation of MarkdownReportsDev
failed, you can also just source this file.
Load packages
# Load proper packages ---------------------------------------------
require(tidyverse); require(cowplot); require(Seurat); require(stringr);
require(tictoc);
library(schex)
# Load custom function libraries ---------------------------------------------
try (source ('https://raw.githubusercontent.com/vertesy/CodeAndRoll/master/CodeAndRoll.R'),silent= F)
try(source('https://raw.githubusercontent.com/vertesy/Seurat.utils/master/00.Load.Seurat.Utils.WEB.R'), silent = T)
try(source("https://raw.githubusercontent.com/vertesy/ggExpressDev/main/ggExpress.functions.R"), silent = T)
# try(source ('https://raw.githubusercontent.com/vertesy/Seurat.multicore/master/00.Load.Seurat3.Multicore.WEB.R'), silent=T) # Not needed I think
# If it fails, see alternative at the end.
require('MarkdownReportsDev')
# If installation of MarkdownReportsDev did not work:
# try (source ('https://raw.githubusercontent.com/vertesy/MarkdownReportsDev/master/R/MarkdownReportsDev.R'),silent= F)
from seurat.pipeline.
Example script for visualization
You can also find it on Seurat.utils
######################################################################
# Visualize Single-cell RNA-seq data using a Seurat object
######################################################################
# source ('Visualize.Gene.expression.v2.R')
# rm(list=ls(all.names = TRUE)); try(dev.off(), silent = T)
# 1. Install function libraries ------------------------
if (FALSE) {
"Run these only once"
install.packages(tidyverse)
install.packages(cowplot)
install.packages(Seurat)
install.packages(tictoc)
install.packages("devtools"); # If you don't have it
require("devtools")
devtools::install_github(repo = "vertesy/MarkdownReportsDev")
}
# 2. Load function libraries ------------------------
# 2a Load proper packages ---------------------------------------------
require(tidyverse); require(cowplot);
require(Seurat);
require(tictoc);
try(library(schex))
require('MarkdownReportsDev')
# If installation of MarkdownReportsDev did not work:
# try (source ('https://raw.githubusercontent.com/vertesy/MarkdownReportsDev/master/R/MarkdownReportsDev.R'),silent= F)
# 2b Load proper packages ---------------------------------------------
try (source ('https://raw.githubusercontent.com/vertesy/CodeAndRoll/master/CodeAndRoll.R'),silent= F)
try(source('https://raw.githubusercontent.com/vertesy/Seurat.utils/master/00.Load.Seurat.Utils.WEB.R'), silent = T)
try(source("https://raw.githubusercontent.com/vertesy/ggExpressDev/main/ggExpress.functions.R"), silent = T)
# 2c Setup ------------------------
"Where do you want to save the output files?"
OutDir = "~/Downloads/scAnalysis"
setup_MarkdownReports(OutDir = OutDir, scriptname = "Visualize.Gene.expression.v2.R")
OutDirOrig = OutDir
# 3. load Seurat object (sc data) -----------------------------------------------------------------
"Where did you save combined.obj.RDS.gz ?"
combined.obj <- readRDS("path/to/combined.obj.RDS.gz")
# 4. create list of genes (all.genes -----------------------------------------------------------------
recall.all.genes()
# 5. search for genes -----------------------------------------------------------------
"Start typing after all.genes$... and hit TAB for autocomplete"
all.genes$SATB2
# 5b. plot genes -----------------------------------------------------------------
qUMAP(feature = "SATB2", obj = combined.obj)
# 6. find.clustering -----------------------------------------------------------------
"Find clustering result: Start typing after combined.ob$... and hit TAB for autocomplete"
combined.obj$integrated_snn_res.0.5
# 6b. plot clusters -----------------------------------------------------------------
clUMAP(ident = , obj = combined.obj)
# Custom gene sets --------------------------------------------------------------------------------
SEGA.old = c("HES5", "ID3", "STMN1", "HMGN2", "SLC6A9", "VCAM1", "S1PR1",
"NOTCH2", "S100A6", "BCAN", "NES", "ATP1A2", "PRDX6", "AGT",
"ID2", "MYCN", "TMSB10", "DBI", "GAD1", "DLX1", "DLX2", "NEUROD1",
"MAP2", "IGFBP5", "ACSL3", "PTMA", "SLC6A11", "SLC6A1", "RPL32",
"RPSA", "SYNPR", "ALDH1L1", "MCM2", "HES1", "FGFR3", "PROM1",
"HOPX", "SPARCL1", "BMPR1B", "H2AFZ", "CCNA2", "CPE", "SLC1A3",
"THBS4", "LIX1", "NREP", "ACSL6", "SPARC", "NPM1", "HNRNPAB",
"ID4", "TFEB", "NR2E1", "GJA1", "FABP7", "MOXD1", "DLL1", "LFNG",
"SP8", "PPIA", "EGFR", "PTN", "RARRES2", "TMSB4X", "TSPAN7",
"DCX", "BMP1", "CLU", "SFRP1", "PABPC1", "MYC", "NTRK2", "TNC",
"RPL12", "CD81", "DKK3", "PAX6", "SLC1A2", "CCND1", "NCAM1",
"PARD3", "CDK1", "HTRA1", "MKI67", "FOXM1", "CCND2", "CD9", "CDCA3",
"ENO2", "SLCO1C1", "LDHB", "LIMA1", "CDK2", "RPL41", "HMGA2",
"ASCL1", "RAN", "TPT1", "ITM2B", "CLDN10", "FOS", "CKB", "NUPR1",
"TUBB3", "TP53", "AURKB", "ALDOC", "GFAP", "LUC7L3", "TOB1",
"SOX9", "RBFOX3", "FASN", "MIB1", "AQP4", "PRNP", "PCNA", "JAG1",
"CST3", "E2F1", "RPL18A", "FXYD1", "DLL3", "APOE", "SMARCB1",
"MIAT", "OLIG2", "GSTM1", "GPR37L1", "BMP6", "CSPG4", "MT1A",
"ETNPPL")
# Markers ----------------------------
ClassicMarkers = c(
"Apical precursor" = "SOX2",
"Stem cells" = "ID4",
"Cycling cells" = "TOP2A",
"IPC" = "EOMES",
"IPC.late" = "TAC3",
"Astro-like" = "S100B",
"Interneurons, (OPC, MSN)" = "DLX6-AS1",
"GABA-ergic inter neuron" = "GAD2",
"General neuronal" = "NEUROD6",
"CTIP2 " = "BCL11B",
"Upper layer " = "SATB2",
"Deep layer neurons" = "FEZF2",
"Weird sc gene" = "MALAT1",
"RPL34" = "RPL34",
"Mito" = "MT-ATP6",
"PDK" = "PDK1",
"SLA" = "SLA",
"IGFBP5" = "IGFBP5"
)
from seurat.pipeline.
Alternative loading of Seurat.utils
and Seurat.multicore
Use these, if the first 2 commands under "2. Load function libraries" above return an error
# Alternaitve for SEURAT.UTILS - You can try to load each script individually ------------------------
if (FALSE) {
"Individual SEURAT.UTILS components"
"You probably dont need all for your work"
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Seurat.update.gene.symbols.HGNC.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Metadata.manipulation.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Plotting.dim.reduction.2D.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Plotting.dim.reduction.3D.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Plotting.filtering.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Plotting.statistics.and.QC.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Read.Write.Save.Load.functions.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Seurat.object.manipulations.etc.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Cluster.Auto-naming.DE.R"))
try(source('https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/MULTI-seq.functions.R'))
try(source('https://raw.githubusercontent.com/vertesy/Seurat.utils/master/Functions/Jaccard.toolkit.R'))
}
# Alternaitve for SEURAT.MULTICORE - You can try to load each script individually ------------------------
if (FALSE) {
" SEURAT.MULTICORE components"
"You probably dont need these at all for your work"
try(source("https://raw.githubusercontent.com/vertesy/Seurat.multicore/master/Seurat3.Multicore.Read.Write.R"))
try(source("https://raw.githubusercontent.com/vertesy/Seurat.multicore/master/Seurat3.Multicore.Generic.Functions.R"))
}
from seurat.pipeline.
Related Issues (9)
- To Do
- AddGOGeneList.manual() used, but now only part of Gruffi's source code HOT 1
- Example
- Needs clean up: `Gene.List.Overlap.R` and `Plot.gene.set.overlaps.R`
- Rename Genes Clarification HOT 1
- Missing iteration index HOT 1
- Gene List plotting is buggy, needs a 'p$' style storage and cleanup.
- Error in STRING.db.auto.cluster.annotation.R HOT 1
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from seurat.pipeline.