Last updated: 2024-07-27
Checks: 7 0
Knit directory: muse/
This reproducible R Markdown analysis was created with workflowr (version 1.7.1). The Checks tab describes the reproducibility checks that were applied when the results were created. The Past versions tab lists the development history.
Great! Since the R Markdown file has been committed to the Git repository, you know the exact version of the code that produced these results.
Great job! The global environment was empty. Objects defined in the global environment can affect the analysis in your R Markdown file in unknown ways. For reproduciblity it’s best to always run the code in an empty environment.
The command set.seed(20200712)
was run prior to running
the code in the R Markdown file. Setting a seed ensures that any results
that rely on randomness, e.g. subsampling or permutations, are
reproducible.
Great job! Recording the operating system, R version, and package versions is critical for reproducibility.
Nice! There were no cached chunks for this analysis, so you can be confident that you successfully produced the results during this run.
Great job! Using relative paths to the files within your workflowr project makes it easier to run your code on other machines.
Great! You are using Git for version control. Tracking code development and connecting the code version to the results is critical for reproducibility.
The results in this page were generated with repository version 00acf0d. See the Past versions tab to see a history of the changes made to the R Markdown and HTML files.
Note that you need to be careful to ensure that all relevant files for
the analysis have been committed to Git prior to generating the results
(you can use wflow_publish
or
wflow_git_commit
). workflowr only checks the R Markdown
file, but you know if there are other scripts or data files that it
depends on. Below is the status of the Git repository when the results
were generated:
Ignored files:
Ignored: .Rhistory
Ignored: .Rproj.user/
Ignored: r_packages_4.3.3/
Ignored: r_packages_4.4.0/
Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.
These are the previous versions of the repository in which changes were
made to the R Markdown (analysis/sradb.Rmd
) and HTML
(docs/sradb.html
) files. If you’ve configured a remote Git
repository (see ?wflow_git_remote
), click on the hyperlinks
in the table below to view the files as they were in that past version.
File | Version | Author | Date | Message |
---|---|---|---|---|
Rmd | 00acf0d | Dave Tang | 2024-07-27 | Join lookup with sample attribute table |
html | 3876763 | Dave Tang | 2024-07-27 | Build site. |
Rmd | e6dba7a | Dave Tang | 2024-07-27 | Format sample attribute |
html | f3646e2 | Dave Tang | 2024-07-27 | Build site. |
Rmd | 3c6da94 | Dave Tang | 2024-07-27 | Get sample information |
html | 7e58cae | Dave Tang | 2024-07-25 | Build site. |
Rmd | a0610af | Dave Tang | 2024-07-25 | Add intro and example query |
html | 1dbd4f3 | Dave Tang | 2024-07-25 | Build site. |
Rmd | 39a6fe8 | Dave Tang | 2024-07-25 | Querying the SRA |
The SRAdb package is a compilation of metadata from NCBI SRA and tools. Specifically:
The Sequence Read Archive (SRA) is the largest public repository of sequencing data from the next generation of sequencing platforms including Roche 454 GS System, Illumina Genome Analyzer, Applied Biosystems SOLiD System, Helicos Heliscope, and others. However, finding data of interest can be challenging using current tools.
SRAdb is an attempt to make access to the metadata associated with submission, study, sample, experiment and run much more feasible. This is accomplished by parsing all the NCBI SRA metadata into a SQLite database that can be stored and queried locally. Fulltext search in the package make querying metadata very flexible and powerful.
fastq and sra files can be downloaded for doing alignment locally. Beside ftp protocol, the SRAdb has funcitons supporting fastp protocol (ascp from Aspera Connect) for faster downloading large data files over long distance. The SQLite database is updated regularly as new data is added to SRA and can be downloaded at will for the most up-to-date metadata.
First download SRAmetadb.sqlite.gz
and gunzip it; the
function getSRAdbFile()
can do this but I recommend
downloading this file externally. The gzipped file is around 6.8G in
size (2024-07-26) and when uncompressed is 138G in size.
sqlfile <- getSRAdbFile(method = "wget")
Download database file externally using wget
.
wget -c https://gbnci.cancer.gov/backup/SRAmetadb.sqlite.gz
Point to the location of downloaded and gunzipped file.
sqlfile <- '/data2/sradb/SRAmetadb.sqlite'
Create a connection for queries. The standard DBI functionality as implemented in RSQLite function dbConnect makes the connection to the database. The dbDisconnect function disconnects the connection.
sra_con <- dbConnect(SQLite(), sqlfile)
Database tables.
sra_tables <- dbListTables(sra_con)
sra_tables
[1] "col_desc" "experiment" "metaInfo" "run"
[5] "sample" "sra" "sra_ft" "sra_ft_content"
[9] "sra_ft_segdir" "sra_ft_segments" "study" "submission"
Study fields.
dbListFields(sra_con, "study")
[1] "study_ID" "study_alias" "study_accession"
[4] "study_title" "study_type" "study_abstract"
[7] "broker_name" "center_name" "center_project_name"
[10] "study_description" "related_studies" "primary_study"
[13] "sra_link" "study_url_link" "xref_link"
[16] "study_entrez_link" "ddbj_link" "ena_link"
[19] "study_attribute" "submission_accession" "sradb_updated"
Sample fields.
dbListFields(sra_con, "sample")
[1] "sample_ID" "sample_alias" "sample_accession"
[4] "broker_name" "center_name" "taxon_id"
[7] "scientific_name" "common_name" "anonymized_name"
[10] "individual_name" "description" "sra_link"
[13] "sample_url_link" "xref_link" "sample_entrez_link"
[16] "ddbj_link" "ena_link" "sample_attribute"
[19] "submission_accession" "sradb_updated"
Query.
rs <- dbGetQuery(sra_con, "select * from study limit 3")
rs[, 1:5]
study_ID study_alias study_accession
1 1 DRP000001 DRP000001
2 2 DRP000002 DRP000002
3 3 DRP000003 DRP000003
study_title
1 Bacillus subtilis subsp. natto BEST195 genome sequencing project
2 Model organism for prokaryotic cell differentiation and development
3 Comprehensive identification and characterization of the nucleosome structure
study_type
1 Whole Genome Sequencing
2 Whole Genome Sequencing
3 Transcriptome Analysis
Query matching specific study accession.
dbGetQuery(sra_con, "select * from study where study_accession == 'DRP000001'")
study_ID study_alias study_accession
1 1 DRP000001 DRP000001
study_title
1 Bacillus subtilis subsp. natto BEST195 genome sequencing project
study_type
1 Whole Genome Sequencing
study_abstract
1 <b><i>Bacillus subtilis</i> subsp. <i>natto</i> BEST195</b>. i>Bacillus subtilis</i> subsp. <i>natto</i> BEST195 was isolated from fermented soybeans and will be used for comparative genome analysis.
broker_name center_name center_project_name
1 <NA> KEIO Bacillus subtilis subsp. natto BEST195
study_description related_studies primary_study sra_link study_url_link
1 <NA> <NA> <NA> <NA> <NA>
xref_link study_entrez_link ddbj_link ena_link
1 pubmed: 20398357 || pubmed: 25329997 <NA> <NA> <NA>
study_attribute submission_accession sradb_updated
1 <NA> DRA000001 2023-12-03 23:10:21
Match a list of study accessions.
rs <- dbGetQuery(sra_con, "select * from study where study_accession in ('DRP000001', 'DRP000003')")
rs[, 1:5]
study_ID study_alias study_accession
1 1 DRP000001 DRP000001
2 3 DRP000003 DRP000003
study_title
1 Bacillus subtilis subsp. natto BEST195 genome sequencing project
2 Comprehensive identification and characterization of the nucleosome structure
study_type
1 Whole Genome Sequencing
2 Transcriptome Analysis
How do we get the SRR accession for SRX510365? (Should be SRR1216135)
exps <- c("SRX510281", "SRX510280", "SRX510279", "SRX510278", "SRX510277", "SRX510276")
lookup <- sraConvert(exps, sra_con = sra_con)
lookup
experiment submission study sample run
1 SRX510276 SRA090948 SRP025982 SRS588793 SRR1216046
2 SRX510277 SRA090948 SRP025982 SRS588794 SRR1216047
3 SRX510278 SRA090948 SRP025982 SRS588795 SRR1216048
4 SRX510279 SRA090948 SRP025982 SRS588796 SRR1216049
5 SRX510280 SRA090948 SRP025982 SRS588798 SRR1216050
6 SRX510281 SRA090948 SRP025982 SRS588797 SRR1216051
Get sample information.
purrr::map_df(lookup$sample, \(x){
dbGetQuery(sra_con, paste0("select * from sample where sample_accession == '", x, "'"))
}) -> sample_info
The required sample information is stored in
sample_attribute
and that needs to be further
formatted.
sample_info |>
as_tibble() |>
select(sample_accession, sample_attribute) |>
tidyr::separate_longer_delim(cols = sample_attribute, delim = " || ") |>
tidyr::separate_wider_delim(cols = sample_attribute, delim = ": ", names = c('key', 'value')) |>
tidyr::pivot_wider(id_cols = sample_accession, names_from = key, values_from = value) -> sample_attribute
Join into one table.
dplyr::inner_join(x = lookup, y = sample_attribute, by = c('sample' = "sample_accession"))
experiment submission study sample run
1 SRX510276 SRA090948 SRP025982 SRS588793 SRR1216046
2 SRX510277 SRA090948 SRP025982 SRS588794 SRR1216047
3 SRX510278 SRA090948 SRP025982 SRS588795 SRR1216048
4 SRX510279 SRA090948 SRP025982 SRS588796 SRR1216049
5 SRX510280 SRA090948 SRP025982 SRS588798 SRR1216050
6 SRX510281 SRA090948 SRP025982 SRS588797 SRR1216051
source_name seqc sample platform
1 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
2 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
3 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
4 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
5 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
6 Human Brain Reference RNA (HBRR) from Ambion B Illumina HiSeq 2000
site library id lane barcode flowcell
1 NYG 4 L03 TAGCTT AC132FACXX
2 NYG 4 L04 TAGCTT AC132FACXX
3 NYG 4 L05 TAGCTT AC132FACXX
4 NYG 4 L06 TAGCTT AC132FACXX
5 NYG 4 L07 TAGCTT AC132FACXX
6 NYG 4 L08 TAGCTT AC132FACXX
Disconnect.
dbDisconnect(conn = sra_con)
sessionInfo()
R version 4.4.0 (2024-04-24)
Platform: x86_64-pc-linux-gnu
Running under: Ubuntu 22.04.4 LTS
Matrix products: default
BLAS: /usr/lib/x86_64-linux-gnu/openblas-pthread/libblas.so.3
LAPACK: /usr/lib/x86_64-linux-gnu/openblas-pthread/libopenblasp-r0.3.20.so; LAPACK version 3.10.0
locale:
[1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C
[3] LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8
[5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8
[7] LC_PAPER=en_US.UTF-8 LC_NAME=C
[9] LC_ADDRESS=C LC_TELEPHONE=C
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C
time zone: Etc/UTC
tzcode source: system (glibc)
attached base packages:
[1] stats graphics grDevices utils datasets methods base
other attached packages:
[1] SRAdb_1.66.0 RCurl_1.98-1.14 graph_1.82.0
[4] BiocGenerics_0.50.0 RSQLite_2.3.7 lubridate_1.9.3
[7] forcats_1.0.0 stringr_1.5.1 dplyr_1.1.4
[10] purrr_1.0.2 readr_2.1.5 tidyr_1.3.1
[13] tibble_3.2.1 ggplot2_3.5.1 tidyverse_2.0.0
[16] workflowr_1.7.1
loaded via a namespace (and not attached):
[1] gtable_0.3.5 xfun_0.44 bslib_0.7.0 processx_3.8.4
[5] Biobase_2.64.0 callr_3.7.6 tzdb_0.4.0 bitops_1.0-7
[9] vctrs_0.6.5 tools_4.4.0 ps_1.7.6 generics_0.1.3
[13] stats4_4.4.0 fansi_1.0.6 blob_1.2.4 pkgconfig_2.0.3
[17] data.table_1.15.4 lifecycle_1.0.4 compiler_4.4.0 git2r_0.33.0
[21] statmod_1.5.0 munsell_0.5.1 getPass_0.2-4 httpuv_1.6.15
[25] htmltools_0.5.8.1 sass_0.4.9 yaml_2.3.8 later_1.3.2
[29] pillar_1.9.0 jquerylib_0.1.4 whisker_0.4.1 limma_3.60.4
[33] cachem_1.1.0 tidyselect_1.2.1 digest_0.6.35 stringi_1.8.4
[37] rprojroot_2.0.4 fastmap_1.2.0 grid_4.4.0 GEOquery_2.72.0
[41] colorspace_2.1-0 cli_3.6.2 magrittr_2.0.3 utf8_1.2.4
[45] withr_3.0.0 scales_1.3.0 promises_1.3.0 bit64_4.0.5
[49] timechange_0.3.0 rmarkdown_2.27 httr_1.4.7 bit_4.0.5
[53] hms_1.1.3 memoise_2.0.1 evaluate_0.24.0 knitr_1.47
[57] rlang_1.1.4 Rcpp_1.0.12 glue_1.7.0 DBI_1.2.3
[61] xml2_1.3.6 rstudioapi_0.16.0 jsonlite_1.8.8 R6_2.5.1
[65] fs_1.6.4