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README.md
<div align="center"> <img src="./logo.png" width="200"> </div> # CellBarcode [![check](https://github.com/wenjie1991/CellBarcode/actions/workflows/r.yml/badge.svg)](https://github.com/wenjie1991/CellBarcode/actions) [![BioC status](http://www.bioconductor.org/shields/build/release/bioc/CellBarcode.svg)](https://bioconductor.org/checkResults/release/bioc-LATEST/CellBarcode) **CellBarcode** is an R package for dealing with **Cellular DNA barcoding** sequencing data. The R package was created by Wenjie SUN, Anne-Marie Lyne, and Leïla Perié at Institut Curie. ## Types of barcodes **CellBarcode** can handle all types of DNA barcodes, provided that: - The barcodes have a pattern that can be matched by a regular expression. - Each barcode is within a single sequencing read. ## What you can do with **CellBarcode** - Perform quality control for the DNA sequence results, and filter the sequences according to their quality metrics. - Identify barcode (and UMI) information in sequencing results. - Performs quality control and deal with the spurious sequences that come from potential PCR & sequence errors. - Provide toolkits to make it easier to manage samples and barcodes with metadata. ## Installing ### Install the development version from GitHub ```r if(!requireNamespace("remotes", quietly = TRUE)) install.packages("remotes") remotes::install_github("wenjie1991/CellBarcode") ``` ## Getting Started Here is an example of a basic workflow: ```r library(CellBarcode) library(magrittr) # The example data is a mix of MEF lines with known barcodes # 2000 reads for each file have been sampled for this test dataset # Data can be accessed here: https://zenodo.org/records/10027002 example_data <- system.file("extdata", "mef_test_data", package = "CellBarcode") fq_files <- dir(example_data, "gz", full=TRUE) # prepare metadata metadata <- stringr::str_split_fixed(basename(fq_files), "_", 10)[, c(4, 6)] metadata <- data.frame(metadata) sample_name <- apply(metadata, 1, paste, collapse = "_") colnames(metadata) = c("cell_number", "replication") rownames(metadata) = sample_name metadata # extract UMI barcode with regular expression bc_obj <- bc_extract( fq_files, pattern = "(.{12})CTCGAGGTCATCGAAGTATCAAG(.+)TAGCAAGCTCGAGAGTAGACCTACT", pattern_type = c("UMI" = 1, "barcode" = 2), sample_name = sample_name, metadata = metadata ) bc_obj # sample subset operation, select 'mixa' bc_sub <- bc_subset(bc_obj, sample=replication == "mixa") bc_sub # filter the barcode, UMI barcode amplicon >= 2 & UMI counts >= 2 bc_sub <- bc_cure_umi(bc_sub, depth = 2) %>% bc_cure_depth(depth = 2) # select barcodes with a white list bc_sub[c("AAGTCCAGTACTATCGTACTA", "AAGTCCAGTACTGTAGCTACTA"), ] # export the barcode counts to data.frame head(bc_2df(bc_sub)) # export the barcode counts to matrix head(bc_2matrix(bc_sub)) ``` ## License [MIT](https://choosealicense.com/licenses/mit/) ## Citation If you use **CellBarcode** in your research, please cite the following paper: [Sun, W. et al. Extracting, filtering and simulating cellular barcodes using CellBarcode tools. Nat Comput Sci 1–16 (2024)](https://www.nature.com/articles/s43588-024-00595-7)