A computational method to reconstruct full-length, paired T cell receptor (TCR) sequences from T lymphocyte single-cell RNA sequence data. TraCeR links T cell specificity with functional response by revealing clonal relationships between cells alongside their transcriptional profiles. TraCeR extracts TCR-derived sequencing reads for each cell by alignment against ‘combinatorial recombinomes’ comprising all possible combinations of V and J segments. Reads are then assembled into contiguous sequences that are analyzed to find full-length, recombined TCR sequences. Importantly, the reconstructed recombinant sequences typically contain nearly the complete length of the TCR V(D)J region and so allow high-confidence discrimination between closely related gene segments. Our method is sensitive, accurate and easy to adapt to any species for which annotated TCR gene sequences are available.
Rebuilds paired full-length B-cell receptor (BCR) sequences. BraCeR is a program which can be used for downstream analyses. This program is able to reconstitute multiple heavy and light chains detected within a target cell as well as to highlight non-productively rearranged chains. This program can also be used as a method for deducing clonal relationships and perform immunoglobulin lineage reconstruction.
CEL-Seq provides its first single-cell, on-chip barcoding method, and we detected gene expression changes accompanying the progression through the cell cycle in mouse fibroblast cells. The pipeline consists of the following steps: (1) demultiplexing: using the barcode from R1 we split R2 reads into their original samples creating a separate file for each sample. Since the unique molecular identifier (UMI) is also read in R1 we extract it and attach it to the R2 read metadata for downstream analysis; (2) mapping: using Bowtie2, we map the reads of the different samples in parallel, cutting the analysis time by roughly the number of available cores; (3) read counting: A modified version of the htseq-count script that supports the identification and elimination of reads sharing the same UMI to generate an accurate molecule count for each feature. We use binomial statistics to convert the number of UMIs into transcript counts. The different steps in the pipeline are wrapped together in a single program with a simple configuration file allowing to control for different run modes.
Processes Chromium single cell 3’ RNA-seq output to align reads, generates gene-cell matrices and performs clustering and gene expression analysis. Cell Ranger combines Chromium-specific algorithms with the widely-used RNA-seq aligner STAR. It is delivered as a single, self-contained tar file that can be unpacked anywhere on the system. The tool includes four pipelines: cellranger mkfastq; cellranger count; cellranger aggr; cellranger reanalyze.
Allows quality control (QC) and analysis components of parallel single cell transcriptome and epigenome data. Dr.seq is a quality control (QC) and analysis pipeline that provides both multifaceted QC reports and cell clustering results. Parallel single cell transcriptome data generated by different technologies can be transformed to the standard input with contained functions. Using relevant commands, the software can also be used to report quality measurements based on four aspects and can generate detailed analysis results for scATAC-seq and Drop-ChIP datasets.
A cloud-based framework designed for multi-sample analysis of transcriptomic data in an efficient and scalable manner. Falco utilises state-of-the-art big data technology of Apache Hadoop and Apache Spark to perform massively parallel alignment, quality control, and feature quantification of single-cell transcriptomic data in Amazon Web Service (AWS) cloud-computing environment. We have evaluated the performance of Falco using two public scRNA-seq datasets and demonstrated Falco's scalability. The result shows Falco performs at least 2.6x faster against a highly optimized single node analysis and a reduction in runtime with increasing number of computing nodes. Falco also allows user to the utilise low-cost spot instances of AWS, providing a 65% reduction in cost of analysis.
Processes raw reads to count tables for RNA-seq data using Unique Molecular Identifiers (UMIs). zUMIs is a pipeline applicable for most experimental designs of RNA-seq data, such as single-nuclei sequencing techniques. This method allows for down sampling of reads before summarizing UMIs per feature, which is recommended for cases of highly different read numbers per sample. zUMIs is flexible with respect to the length and sequences of the barcodes (BCs) and UMIs, making it compatible with a large number of protocols.
Addresses the lack of a comprehensive workflow for processing sequencing data from 3 prime end protocols. scPipe can deal with both unique molecular identifiers (UMIs) and sample barcodes, map reads to the genome and summarizes these results into gene-level counts. It implements a simple outlier-based method for discovering low quality cells and possible doublets to remove from further analysis.
Allows users to transform raw data from dropSeq/scrbSeq experiment to the final count matrix with QC plots. dropSeqPipe is an open source application that can perform five different tasks: (i) generate fastqc reports of the input data, (ii) obtain the final file for the aligned sorted data, (iii) produce plots based on pre-processing and alignement, (iv) create species plot, and (v) extract the expression data.
Assists in processing single cell RNA-seq (scRNA-seq) FASTQ reads generated by CEL-Seq and CEL-Seq2 protocols. SCRUFF permits users to demultiplexe scRNA-seq FASTQ files, align reads to reference genome using Rsubread, and generate unique molecular identifier (UMI) filtered count matrix. This filtering facilitator is available as an R package.
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