## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # library(ORFik) # This package # conf <- config.exper(experiment = "CHALMERS_Yeast", # Name # assembly = "Yeast_SacCer3", # Reference folder # type = c("RNA-seq")) # fastq and bam type ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # # info <- download.SRA.metadata("SRP012047", outdir = conf["fastq RNA-seq"]) # # Let's take 2 first runs in this experiment: # info <- info[1:2,] # # 18 MB, ~ 40 sec download time -> # download.SRA(info, conf["fastq RNA-seq"], subset = 50000) # # 1.6 GB, ~ 100 sec download time (faster download) -> # # download.SRA(info, conf["fastq RNA-seq"]) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # organism <- info$ScientificName[1] # is_paired_end <- all(info$LibraryLayout == "PAIRED") ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # annotation <- getGenomeAndAnnotation( # organism = organism, # output.dir = conf["ref"], # assembly_type = "toplevel" # ) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # index <- STAR.index(annotation) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # alignment <- # STAR.align.folder(conf["fastq RNA-seq"], conf["bam RNA-seq"], index, # paired.end = is_paired_end, # steps = "tr-ge", # (trim needed: adapters found, then genome) # adapter.sequence = "auto", # max.cpus = 30, trim.front = 3, min.length = 20) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # index <- STAR.index(annotation, max.ram = 20, SAsparse = 2) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # STAR.align.folder(conf["fastq RNA-seq"], conf["bam RNA-seq"], index, # max.cpus = 12) # Reduce cores to 12 usually works for most systems ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # txdb_file <- paste0(annotation["gtf"], ".db") # Get txdb file, not raw gtf # fa <- annotation["genome"] # create.experiment(exper = "yeast_exp_RNA", # dir = paste0(conf["bam RNA-seq"], "/aligned/"), # txdb = txdb_file, fa = fa, # organism = organism, # viewTemplate = FALSE, # pairedEndBam = is_paired_end # True/False per bam file # ) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # df <- read.experiment("yeast_exp_RNA") ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # QCreport(df) ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # remove.experiments(df) # Remove loaded libraries # convertLibs(df, type = "ofst") ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # remove.experiments(df) # convertLibs(df, type = "wig") ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # remove.experiments(df) # outputLibs(df, type = "ofst") ## ----eval = FALSE, echo = TRUE, message = FALSE------------------------------- # mrna <- countTable(df, region = "mrna", type = "fpkm") # cds <- countTable(df, region = "cds", type = "fpkm") # ratio <- cds / mrna