Created
March 18, 2016 18:47
-
-
Save pimentel/5d13acfaa2074cb761c5 to your computer and use it in GitHub Desktop.
very simple sleuth/DESeq2 comparison at transcript level
This file contains hidden or bidirectional Unicode text that may be interpreted or compiled differently than what appears below. To review, open the file in an editor that reveals hidden Unicode characters.
Learn more about bidirectional Unicode characters
suppressPackageStartupMessages({ | |
library('Biobase') | |
library('DESeq2') | |
library('sleuth') | |
}) | |
base_dir <- "~/Downloads/cuffdiff2_data_kallisto_results" | |
sample_id <- dir(file.path(base_dir,"results")) | |
kal_dirs <- sapply(sample_id, function(id) file.path(base_dir, "results", id, "kallisto")) | |
s2c <- read.table(file.path(base_dir, "hiseq_info.txt"), header = TRUE, stringsAsFactors=FALSE) | |
s2c <- dplyr::select(s2c, sample = run_accession, condition) | |
s2c <- dplyr::mutate(s2c, path = kal_dirs) | |
mart <- biomaRt::useMart(biomart = "ENSEMBL_MART_ENSEMBL", | |
dataset = "hsapiens_gene_ensembl", | |
host = "may2015.archive.ensembl.org") | |
t2g <- biomaRt::getBM(attributes = c("ensembl_transcript_id", "ensembl_gene_id", | |
"external_gene_name"), mart = mart) | |
t2g <- dplyr::rename(t2g, target_id = ensembl_transcript_id, | |
ens_gene = ensembl_gene_id, ext_gene = external_gene_name) | |
suppressMessages(so <- sleuth_prep(s2c, ~ condition, target_mapping = t2g)) | |
so <- sleuth_fit(so) | |
so <- sleuth_wt(so, 'conditionscramble') | |
sr <- sleuth_results(so, 'conditionscramble') | |
sr_sig <- dplyr::filter(sr, qval <= 0.01) | |
### | |
# DESeq2 | |
### | |
obs_raw <- sleuth:::spread_abundance_by(so$obs_raw, "est_counts") | |
obs_raw <- round(obs_raw) | |
mode(obs_raw) <- 'integer' | |
s2c <- as.data.frame(s2c) | |
rownames(s2c) <- s2c$sample | |
cds <- ExpressionSet(obs_raw, AnnotatedDataFrame(s2c)) | |
dds <- DESeqDataSetFromMatrix(exprs(cds), DataFrame(pData(cds)), ~ condition) | |
dds <- dds[ rowSums(counts(dds)) > 1, ] | |
dds <- DESeq(dds,quiet=TRUE) | |
ds_res <- results(dds) | |
ds_res$target_id <- rownames(ds_res) | |
ds_res <- as.data.frame(ds_res) | |
ds_sig <- dplyr::filter(ds_res, padj <= 0.01) | |
### | |
# comparison | |
### | |
nrow(sr_sig) | |
nrow(ds_sig) | |
length(intersect(sr_sig$target_id, ds_sig$target_id)) |
Sign up for free
to join this conversation on GitHub.
Already have an account?
Sign in to comment