diff --git a/README.md b/README.md index 96d6cf043..378fdc8a9 100644 --- a/README.md +++ b/README.md @@ -240,6 +240,9 @@ The folder is structured in the following way: Any nonspecific-pangenome file is - test_10x_10k_pbmc_cmo_cmo_fastq_\{1,2\}_gz: 3' V3 Cell Multiplexing FASTQs from 10k PBMC data with Cell Multiplexing - test_10x_10k_pbmc_cmo_gex\{1,2\}_fastq_\{1,2\}_gz: 3' V3 gene expression FASTQs from 10k PBMC data with Cell Multiplexing - test_10x_10k_pbmc_cmo_feature_ref_csv: Feature Barcode reference for the 10k PBMC data with Cell Multiplexing + - test_10x_60k_pbmc_cmo_tcr_gex_fastq_\{1,2\}_gz: 5' V2 HT gene expression FASTQs from 60k PBMC data with Cell Multiplexing + - test_10x_60k_pbmc_cmo_tcr_ab_fastq_\{1,2\}_gz: 5' V2 HT Feature Barcode & Cell Multiplexing FASTQs from 60k PBMC data (files contain both) + - test_10x_60k_pbmc_cmo_tcr_tcr_fastq_\{1,2\}_gz: 5' V2 HT T-cell V(D)J FASTQs from 60k PBMC data with Cell Multiplexing - test_10x_5k_cmvpos_tcells_ab_fastq_\{1,2\}_gz: Antibody Capture FASTQs from the 5k CMV+ T-cell dataset - test_10x_5k_cmvpos_tcells_gex1_fastq_\{1,2\}_gz: Gene expression FASTQs from the 5k CMV+ T-cell dataset - test_10x_5k_cmvpos_tcells_vdj_fastq_\{1,2\}_gz: V(D)J FASTQs from the 5k CMV+ T-cell dataset diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor1_config.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor1_config.csv new file mode 100644 index 000000000..645265147 --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor1_config.csv @@ -0,0 +1,17 @@ +[gene-expression] +reference,working-directory/refdata-gex-GRCh38-2020-A/ +force-cells,11638 +check-library-compatibility,false + +[feature] +reference,working-directory/final-analysis/feature_ref.csv + +[vdj] +reference,working-directory/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,working-directory/bamtofastq/Donor1_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,working-directory/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,working-directory/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,working-directory/fastqs/bcr_fastq,VDJ-B diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor2_config.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor2_config.csv new file mode 100644 index 000000000..644b22141 --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor2_config.csv @@ -0,0 +1,17 @@ +[gene-expression] +reference,working-directory/refdata-gex-GRCh38-2020-A/ +force-cells,8873 +check-library-compatibility,false + +[feature] +reference,working-directory/final-analysis/feature_ref.csv + +[vdj] +reference,working-directory/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,working-directory/bamtofastq/Donor2_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,working-directory/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,working-directory/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,working-directory/fastqs/bcr_fastq,VDJ-B diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor3_config.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor3_config.csv new file mode 100644 index 000000000..367371d1e --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor3_config.csv @@ -0,0 +1,17 @@ +[gene-expression] +reference,working-directory/refdata-gex-GRCh38-2020-A/ +force-cells,9024 +check-library-compatibility,false + +[feature] +reference,working-directory/final-analysis/feature_ref.csv + +[vdj] +reference,working-directory/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,working-directory/bamtofastq/Donor1_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,working-directory/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,working-directory/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,working-directory/fastqs/bcr_fastq,VDJ-B diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor4_config.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor4_config.csv new file mode 100644 index 000000000..9fbd8d96b --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/Donor4_config.csv @@ -0,0 +1,17 @@ +[gene-expression] +reference,path/to/cellranger/prebuilt/human/refdata-gex-GRCh38-2020-A/ +force-cells,7032 +check-library-compatibility,false + +[feature] +reference,/path/to/feature_ref.csv + +[vdj] +reference,/path/to/vdj/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,/path/to/bamtofastq_outs_v1.4/Donor4_ALLpatient/PBMC_60k_10x_4tags_Rep1_step1_v7_0_1_HMW77DSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,/path/to/Ab-fastqs,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,/path/to/Ab-fastqs/TCR,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,/path/to/Ab-fastqs/BCR/,VDJ-B diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/README.md b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/README.md new file mode 100644 index 000000000..2dce5701c --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/README.md @@ -0,0 +1,209 @@ +# 60k Human PBMCs Multiplexed, 4 CMOs + +Peripheral blood mononuclear cells (PBMCs). Samples originate from two healthy donors (Donor 1 & 2), and from two patients diagnosed with Acute Lymphocytic Leukemia (Donor 3 & 4). Samples from donor 1,2, and 4 consist of peripheral blood mononuclear cells (PBMCs), while the sample from donor 3 consists of Bone Marrow Mononuclear Cells (BMMCs). +Samples stained with TotalSeq-C antibodies both for cell surface protein (CSP) feature barcoding, but also for cell hashing, ie mimicing 3' cell multiplexing oligos (CMO). +Samples were pooled in a 1:1 ratio. + +Libraries were prepared following the Single Cell V(D)J Reagent Kits User Guide CG000424 (5' v2 HT) and sequenced on Illumina NovaSeq 6000. + +## Download original input data +Data is available from 10x: +[Demultiplexing 5’ Immune Profiling Libraries Pooled with Hashtags](https://www.10xgenomics.com/datasets/5-hashing-example-with-tabs-2-standard) + +```bash +# Input Files +curl -O https://s3-us-west-2.amazonaws.com/10x.files/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_gex_fastq.tar +curl -O https://s3-us-west-2.amazonaws.com/10x.files/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_ab_fastq.tar +curl -O https://s3-us-west-2.amazonaws.com/10x.files/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_tcr_fastq.tar +curl -O https://cf.10xgenomics.com/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_bcr_fastq.tar +curl -O https://cf.10xgenomics.com/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_cmo-set.csv +curl -O https://cf.10xgenomics.com/samples/cell-vdj/7.0.0/5p_hashing_demux/5p_hashing_demux_config.csv +``` + +## Original `cellranger multi` configs +Since this data consist of both multiplexed samples and immune profiling data, data must be processed as follows: + +1. Demultiplex by sample +2. Convert BAM to FASTQ per sample +3. Run cellranger for each sample + +See [this](https://www.10xgenomics.com/analysis-guides/demultiplexing-and-analyzing-5’-immune-profiling-libraries-pooled-with-hashtags) analysis guide. + +### Sample demultiplexing config (step 1) +This config is to demultiplex by sample. This step requires GEX and CMO data. + +```bash +[gene-expression] +reference,/path/to/cellranger/prebuilt/human/refdata-gex-GRCh38-2020-A/ +cmo-set,/path/to/wrkdir/demultiplexing/5p_hashing_demux_cmo-set.csv + +[libraries] +fastq_id,fastqs,feature_types +PBMC-ALL_60k_universal_HashAB1-4_BL_4tags_Rep1_gex,Path/to/GEX/FASTQs,Gene Expression +PBMC-ALL_60k_universal_HashAB1-4_BL_4tags_Rep1_ab,Path/to/AB/FASTQs,Multiplexing Capture + +[samples] +sample_id,cmo_ids +Donor1_healthy,Hash-tag1 +Donor2_healthy,Hash-tag2 +Donor3_ALLpatient,Hash-tag3 +Donor4_ALLpatient,Hash-tag4 +``` + +### Convert BAM to FASTQ (step 2) +For each sample output convert the GEX BAM file to FASTQ. +This step requires the 10x provided bamtofastq binaries which is bundled with cellranger. +Add `bamtofastq`to your `$PATH` like so: + +```bash +# Command to change to the directory where the Cell Ranger executable file lives and put it in your $PATH: +cd /path/to/cellranger-7.0.0/ +export PATH=${PWD}:$PATH + +# Command to put other tools bundled with Cell Ranger in your path: +source /path/to/cellranger-7.0.0/sourceme.bash +``` + +Run `bamtofastq` per sample: +```bash +mkdir bamtofastq + +bamtofastq /path/to/wrkdir/demultiplexing/demultiplexed_samples/outs/per_sample_outs/Donor1_healthy/count/sample_alignments.bam /path/to/wrkdir/bamtofastq/Donor1_healthy +bamtofastq /path/to/wrkdir/demultiplexing/demultiplexed_samples/outs/per_sample_outs/Donor2_healthy/count/sample_alignments.bam /path/to/wrkdir/bamtofastq/Donor2_healthy +bamtofastq /path/to/wrkdir/demultiplexing/demultiplexed_samples/outs/per_sample_outs/Donor3_ALLpatient/count/sample_alignments.bam /path/to/wrkdir/bamtofastq/Donor3_ALLpatient +bamtofastq /path/to/wrkdir/demultiplexing/demultiplexed_samples/outs/per_sample_outs/Donor4_ALLpatient/count/sample_alignments.bam /path/to/wrkdir/bamtofastq/Donor4_ALLpatient +``` + +Expected output: +```bash +$ ls /path/to/wrkdir/bamtofastq/Donor1_healthy + +demultiplexed_samples_0_1_HW7KMDSX3/ demultiplexed_samples_1_1_HW7WGDSX3/ +``` + +Please note that the order of the libraries depends on the order that the libraries are listed in 5p_hashing_demux_config.csv (Step 1). +The GEX library is listed before Multiplexing Capture, and therefore GEX data is found in `demultiplexed_samples_0_1_HW7KMDSX3/`. + +### Immune profiling per sample (step 3) +This step must be run for each sample. + +```bash +mkdir final-analysis +``` + +Below are listed configs for each sample. + +1. Donor +```bash +[gene-expression] +reference,wrkdir/refdata-gex-GRCh38-2020-A/ +force-cells,11638 +check-library-compatibility,false + +[feature] +reference,wrkdir/final-analysis/feature_ref.csv + +[vdj] +reference,wrkdir/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,wrkdir/bamtofastq/Donor1_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,wrkdir/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,wrkdir/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,wrkdir/fastqs/bcr_fastq,VDJ-B +``` + +2. Donor 2 +```bash +[gene-expression] +reference,wrkdir/refdata-gex-GRCh38-2020-A/ +force-cells,8873 +check-library-compatibility,false + +[feature] +reference,wrkdir/final-analysis/feature_ref.csv + +[vdj] +reference,wrkdir/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,wrkdir/bamtofastq/Donor2_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,wrkdir/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,wrkdir/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,wrkdir/fastqs/bcr_fastq,VDJ-B +``` + +3. Donor 3 +```bash +[gene-expression] +reference,wrkdir/refdata-gex-GRCh38-2020-A/ +force-cells,9024 +check-library-compatibility,false + +[feature] +reference,wrkdir/final-analysis/feature_ref.csv + +[vdj] +reference,wrkdir/final-analysis/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,wrkdir/bamtofastq/Donor1_healthy/demultiplexed_samples_0_1_HW7KMDSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,wrkdir/fastqs/ab_fastq,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,wrkdir/fastqs/tcr_fastq,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,wrkdir/fastqs/bcr_fastq,VDJ-B +``` + +4. Donor 4 +```bash +[gene-expression] +reference,path/to/cellranger/prebuilt/human/refdata-gex-GRCh38-2020-A/ +force-cells,7032 +check-library-compatibility,false + +[feature] +reference,/path/to/feature_ref.csv + +[vdj] +reference,/path/to/vdj/refdata-cellranger-vdj-GRCh38-alts-ensembl-7.0.0 + +[libraries] +fastq_id,fastqs,feature_types +bamtofastq,/path/to/bamtofastq_outs_v1.4/Donor4_ALLpatient/PBMC_60k_10x_4tags_Rep1_step1_v7_0_1_HMW77DSX3,Gene Expression +PBMC_60k_10x_4tags_Rep1_Ab,/path/to/Ab-fastqs,Antibody Capture +PBMC_60k_10x_4tags_Rep1_TCR,/path/to/Ab-fastqs/TCR,VDJ-T +PBMC_60k_10x_4tags_Rep1_BCR,/path/to/Ab-fastqs/BCR/,VDJ-B +``` + +Run cellranger multi: +```bash +cellranger multi --id=Donor1-final --csv=Donor1_config.csv +cellranger multi --id=Donor2-final --csv=Donor2_config.csv +cellranger multi --id=Donor3-final --csv=Donor3_config.csv +cellranger multi --id=Donor4-final --csv=Donor4_config.csv +``` + +## Cell Multiplexing Data + +The cell multiplexing libraries require more generous subsampling to yield sufficient reads for successful processing. +Test data use the first 3,500,000 lines of each of the lane 1 FASTQs. + +```bash +#!/usr/bin/env bash + +ft_type=$1 + +old="raw/fastq/${ft_type}/${ft_type}-PBMC-Control-1" +for of in "${old}_"*.fastq.gz; do + new="raw_sub/fastqs/${ft_type}/subsampled_sc5p_PBMC_60k_${ft_type}" + nf=${of/$old/$new} + echo $of + echo $nf + + zcat "$of" | \ + head -n 3500000 | \ + gzip - > $nf +done +``` diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/cmo-set.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/cmo-set.csv new file mode 100644 index 000000000..eb38fa578 --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/cmo-set.csv @@ -0,0 +1,5 @@ +id,name,read,pattern,sequence,feature_type +Hash-tag1,Hash-tag1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTCAACTCTTTAGCG,Multiplexing Capture 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b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/fastqs/TCR/subsampled_sc5p_PBMC_60k_TCR_S1_L001_R1_001.fastq.gz new file mode 100644 index 000000000..c2e3692d9 Binary files /dev/null and b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/fastqs/TCR/subsampled_sc5p_PBMC_60k_TCR_S1_L001_R1_001.fastq.gz differ diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/fastqs/TCR/subsampled_sc5p_PBMC_60k_TCR_S1_L001_R2_001.fastq.gz b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/fastqs/TCR/subsampled_sc5p_PBMC_60k_TCR_S1_L001_R2_001.fastq.gz new file mode 100644 index 000000000..a54c71594 Binary files /dev/null and b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/fastqs/TCR/subsampled_sc5p_PBMC_60k_TCR_S1_L001_R2_001.fastq.gz differ diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/feature-ref.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/feature-ref.csv new file mode 100644 index 000000000..1ac99c2f1 --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/feature-ref.csv @@ -0,0 +1,138 @@ +id,name,read,pattern,sequence,feature_type +CD86,anti-human_CD86,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTCTTTGTCAGTGCA,Antibody Capture +CD274,anti-human_CD274_B7-H1_PD-L1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTTGTCCGACAATAC,Antibody Capture +CD270,anti-human_CD270_HVEM_TR2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGATAGAAACAGACC,Antibody Capture +CD155,anti-human_CD155_PVR,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATCACATCGTTGCCA,Antibody Capture +CD112,anti-human_CD112_Nectin-2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AACCTTCCGTCTAAG,Antibody Capture +CD47,anti-human_CD47,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCATTCTGTCACCTA,Antibody Capture +CD48,anti-human_CD48,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTACGACGTAGAAGA,Antibody Capture +CD40,anti-human_CD40,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTCAGATGGAGTATG,Antibody Capture +CD154,anti-human_CD154,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCTAGATAGATGCAA,Antibody Capture +CD52,anti-human_CD52,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTTTGTACGAGCAAA,Antibody Capture +CD3,anti-human_CD3,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTCATTGTAACTCCT,Antibody Capture +CD8,anti-human_CD8,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCGCAACTTGATGAT,Antibody Capture +CD56,anti-human_CD56,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCCTTTCCTGATAGG,Antibody Capture +CD19,anti-human_CD19,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTGGGCAATTACTCG,Antibody Capture +CD33,anti-human_CD33,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TAACTCAGGGCCTAT,Antibody Capture +CD11c,anti-human_CD11c,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TACGCCTATAACTTG,Antibody Capture +HLA-A_B_C,anti-human_HLA-A_B_C,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TATGCGAGGCTTATC,Antibody Capture +CD45RA,anti-human_CD45RA,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCAATCCTTCCGCTT,Antibody Capture +CD123,anti-human_CD123,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTTCACTCTGTCAGG,Antibody Capture +CD7,anti-human_CD7,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGGATTCCCGGACTT,Antibody Capture +CD105,anti-human_CD105,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATCGTCGAGAGCTAG,Antibody Capture +CD49f,anti-human_mouse_CD49f,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTCCGAGGATGATCT,Antibody Capture +CD194,anti-human_CD194_CCR4,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGCTTACCTGCACGA,Antibody Capture +CD4,anti-human_CD4,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGTTCCCGCTCAACT,Antibody Capture +CD44,anti-mouse_human_CD44,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGGCTTCAGGTCCTA,Antibody Capture +CD14,anti-human_CD14,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCTCAGACCTCCGTA,Antibody Capture +CD16,anti-human_CD16,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AAGTTCACTCTTTGC,Antibody Capture +CD25,anti-human_CD25,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTTGTCCTGTACGCC,Antibody Capture +CD45RO,anti-human_CD45RO,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTCCGAATCATGTTG,Antibody Capture +CD279,anti-human_CD279,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACAGCGCCGTATTTA,Antibody Capture +TIGIT,anti-human_TIGIT_VSTM3,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTGCTTACCGCCAGA,Antibody Capture +Mouse_IgG1_k_isotype_Ctrl,Mouse_IgG1_k_isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCCGGACGACATTAA,Antibody Capture +Mouse_IgG2a_k_isotype_Ctrl,Mouse_IgG2a_k_isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTCCTACCTAAACTG,Antibody Capture +Mouse_IgG2b_k_isotype_Ctrl,Mouse_IgG2b_k_isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATATGTATCACGCGA,Antibody Capture +Rat_IgG2b_k_isotype_Ctrl,Rat_IgG2b_k_Isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GATTCTTGACGACCT,Antibody Capture +CD20,anti-human_CD20,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTCTGGGTCCCTAGA,Antibody Capture +CD335,anti-human_CD335_NKp46,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACAATTTGAACAGCG,Antibody Capture +CD31,anti-human_CD31,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACCTTTATGCCACGG,Antibody Capture +CD146,anti-human_CD146,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCTTGGATAACATCA,Antibody Capture +IgM,anti-human_IgM,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TAGCGAGCCCGTATA,Antibody Capture +CD5,anti-human_CD5,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CATTAACGGGATGCC,Antibody Capture +CD183,anti-human_CD183_CXCR3,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCGATGGTAGATTAT,Antibody Capture +CD195,anti-human_CD195_CCR5,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCAAAGTAAGAGCCA,Antibody Capture +CD32,anti-human_CD32,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCTTCCGAATTACCG,Antibody Capture +CD196,anti-human_CD196_CCR6,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GATCCCTTTGTCACT,Antibody Capture +CD185,anti-human_CD185_CXCR5,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AATTCAACCGTCGCC,Antibody Capture +CD103,anti-human_CD103_Integrin_aE,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GACCTCATTGTGAAT,Antibody Capture +CD69,anti-human_CD69,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTCTCTTGGCTTAAA,Antibody Capture +CD62L,anti-human_CD62L,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTCCCTGCAACTTGA,Antibody Capture +CD161,anti-human_CD161,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTACGCAGTCCTTCT,Antibody Capture +CD152,anti-human_CD152_CTLA-4,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATGGTTCACGTAATC,Antibody Capture +CD223,anti-human_CD223_LAG-3,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CATTTGTCTGCCGGT,Antibody Capture +KLRG1,anti-human_KLRG1_MAFA,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTTATTTCCTGCCCT,Antibody Capture +CD27,anti-human_CD27,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCACTCCTGCATGTA,Antibody Capture +CD107a,anti-human_CD107a_LAMP-1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CAGCCCACTGCAATA,Antibody Capture +CD95,anti-human_CD95_Fas,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCAGCTCATTAGAGC,Antibody Capture +CD134,anti-human_CD134_OX40,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AACCCACCGTTGTTA,Antibody Capture +HLA-DR,anti-human_HLA-DR,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AATAGCGAGCAAGTA,Antibody Capture +CD1c,anti-human_CD1c,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GAGCTACTTCACTCG,Antibody Capture +CD11b,anti-human_CD11b,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GACAAGTGATCTGCA,Antibody Capture +CD64,anti-human_CD64,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AAGTATGCCCTACGA,Antibody Capture +CD141,anti-human_CD141_Thrombomodulin,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GGATAACCGCGCTTT,Antibody Capture +CD1d,anti-human_CD1d,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCGAGTCGCTTATCA,Antibody Capture +CD314,anti-human_CD314_NKG2D,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CGTGTTTGTTCCTCA,Antibody Capture +CD35,anti-human_CD35,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACTTCCGTCGATCTT,Antibody Capture +CD57,anti-human_CD57_Recombinant,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AACTCCCTATGGAGG,Antibody Capture +CD272,anti-human_CD272_BTLA,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTTATTGGACTAAGG,Antibody Capture +CD278,anti-human_mouse_rat_CD278_ICOS,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CGCGCACCCATTAAA,Antibody Capture +CD58,anti-human_CD58_LFA-3,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTTCCTATGGACGAC,Antibody Capture +CD39,anti-human_CD39,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTACCTGGTATCCGT,Antibody Capture +CX3CR1,anti-human_CX3CR1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGTATCGTCTCTGGG,Antibody Capture +CD24,anti-human_CD24,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGATTCCTTCGTGTT,Antibody Capture +CD21,anti-human_CD21,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AACCTAGTAGTTCGG,Antibody Capture +CD11a,anti-human_CD11a,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TATATCCTTGTGAGC,Antibody Capture +CD79b,anti-human_CD79b_Ig,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATTCTTCAACCGAAG,Antibody Capture +CD244,anti-human_CD244_2B4,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCGCTTGGATGGTAG,Antibody Capture +CD169,anti-human_CD169_Sialoadhesin_Siglec-1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TACTCAGCGTGTTTG,Antibody Capture +integrin_b7,anti-human_mouse_integrin_b7,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCCTTGGATGTACCG,Antibody Capture +CD268,anti-human_CD268_BAFF-R,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CGAAGTCGATCCGTA,Antibody Capture +CD42b,anti-human_CD42b,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCCTAGTACCGAAGT,Antibody Capture +CD54,anti-human_CD54,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTGATAGACTTGAGT,Antibody Capture +CD62P,anti-human_CD62P_P-Selectin,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCTTCCGTATCCCTT,Antibody Capture +CD119,anti-human_CD119_IFN-y_R_a_chain,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGTGTATTCCCTTGT,Antibody Capture +TCRa_b,anti-human_TCR_a_or_b,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CGTAACGTAGAGCGA,Antibody Capture +Rat_IgG1_k_isotype_Ctrl,Rat_IgG1_k_isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ATCAGATGCCCTCAT,Antibody Capture +Rat_IgG2a_k_isotype_Ctrl,Rat_IgG2a_k_Isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AAGTCAGGTTCGTTT,Antibody Capture +Armenian_Hamster_IgG_Isotype_Ctrl,Armenian_Hamster_IgG_Isotype_Ctrl,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCTGTCATTAAGACT,Antibody Capture +CD122,anti-human_CD122_IL-2Rb,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCATTTCCTCCGATT,Antibody Capture +CD267,anti-human_CD267_TACI,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGTGATGGAGCGAAC,Antibody Capture +FceRIa,anti-human_FceRIa,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTCGTTTCCGTATCG,Antibody Capture +CD41,anti-human_CD41,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACGTTGTGGCCTTGT,Antibody Capture +CD137,anti-human_CD137_4-1BB,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CAGTAAGTTCGGGAC,Antibody Capture +CD163,anti-human_CD163,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCTTCTCCTTCCTTA,Antibody Capture +CD83,anti-human_CD83,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCACTCATTTCCGGT,Antibody Capture +CD124,anti-human_CD124_IL-4R,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCGTCCTGATAGATG,Antibody Capture +CD13,anti-human_CD13,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTTCAACGCCCTTTC,Antibody Capture +CD2,anti-human_CD2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TACGATTTGTCAGGG,Antibody Capture +CD226,anti-human_CD226_DNAM-1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCTCAGTGTTTGTGG,Antibody Capture +CD29,anti-human_CD29,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTATTCCCTCAGTCA,Antibody Capture +CD303,anti-human_CD303_BDCA-2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GAGATGTCCGAATTT,Antibody Capture +CD49b,anti-human_CD49b,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCTTTCTTCAGTATG,Antibody Capture +CD81,anti-human_CD81_TAPA-1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTATCCTTCCTTGGC,Antibody Capture +IgD,anti-human_IgD,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CAGTCTCCGTAGAGT,Antibody Capture +CD18,anti-human_CD18,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TATTGGGACACTTCT,Antibody Capture +CD28,anti-human_CD28,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGAGAACGACCCTAA,Antibody Capture +CD38,anti-human_CD38,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGTACCCGCTTGTGA,Antibody Capture +CD127,anti-human_CD127_IL-7Ra,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GTGTGTTGTCCTATG,Antibody Capture +CD45,anti-human_CD45,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TGCAATTACCCGGAT,Antibody Capture +CD22,anti-human_CD22,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GGGTTGTTGTCTTTG,Antibody Capture +CD71,anti-human_CD71,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCGTGTTCCTCATTA,Antibody Capture +CD26,anti-human_CD26,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GGTGGCTAGATAATG,Antibody Capture +CD36,anti-human_CD36,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TTCTTTGCCTTGCCA,Antibody Capture +CD158,anti-human_CD158_KIR2DL1_S1_S3_S5,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TATCAACCAACGCTT,Antibody Capture +CD49a,anti-human_CD49a,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACTGATGGACTCAGA,Antibody Capture +CD49d,anti-human_CD49d,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCATTCAACTTCCGG,Antibody Capture +CD73,anti-human_CD73_Ecto-5-nucleotidase,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CAGTTCCTCAGTTCG,Antibody Capture +TCRV_7-2,anti-human_TCR_V_7-2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TACGAGCAGTATTCA,Antibody Capture +TCRV_2,anti-human_TCR_V_2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCAGTCAGATGGTAT,Antibody Capture +LOX-1,anti-human_LOX-1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACCCTTTACCGAATA,Antibody Capture +CD158b,anti-human_CD158b_KIR2DL2_L3_NKAT2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GACCCGTAGTTTGAT,Antibody Capture +CD158e1,anti-human_CD158e1_KIR3DL1_NKB1,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GGACGCTTTCCTTGA,Antibody Capture +CD319,anti-human_CD319_CRACC,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGTATGCCATGTCTT,Antibody Capture +CD99,anti-human_CD99,R2,^NNNNNNNNNN(BC)NNNNNNNNN,ACCCGTCCCTAAGAA,Antibody Capture +CLEC12A,anti-human_CLEC12A,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CATTAGAGTCTGCCA,Antibody Capture +CD352,anti-human_CD352_NTB-A,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGTTTCCACTCAGGC,Antibody Capture +CD94,anti-human_CD94,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTTTCCGGTCCTACA,Antibody Capture +Ig_light_chain_k,anti-human_Ig_light_chain_k,R2,^NNNNNNNNNN(BC)NNNNNNNNN,AGCTCAGCCAGTATG,Antibody Capture +CD85j,anti-human_CD85j_ILT2,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CCTTGTGAGGCTATG,Antibody Capture +CD23,anti-human_CD23,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCTGTATAACCGTCT,Antibody Capture +Ig_light_chain_g,anti-human_Ig_light_chain_gamma,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CAGCCAGTAAGTCAC,Antibody Capture +CD328,anti-human_CD328_Siglec-7,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTTAGCATTTCACTG,Antibody Capture +GPR56,anti-human_GPR56,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCCTAGTTTCCGTTT,Antibody Capture +HLA-E,anti-human_HLA-E,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GAGTCGAGAAATCAT,Antibody Capture +CD82,anti-human_CD82,R2,^NNNNNNNNNN(BC)NNNNNNNNN,TCCCACTTCCGCTTT,Antibody Capture +CD101,anti-human_CD101_BB27,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTACTTCCCTGTCAA,Antibody Capture +CD88,anti-human_CD88_C5aR,R2,^NNNNNNNNNN(BC)NNNNNNNNN,GCCGCATGAGAAACA,Antibody Capture +CD224,anti-human_CD224,R2,^NNNNNNNNNN(BC)NNNNNNNNN,CTGATGAGATGTCAG,Antibody Capture diff --git a/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/sample_demultiplexing_config.csv b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/sample_demultiplexing_config.csv new file mode 100644 index 000000000..cb622a4fa --- /dev/null +++ b/data/genomics/homo_sapiens/10xgenomics/cellranger/60k_pbmc_cmo_tcr/sample_demultiplexing_config.csv @@ -0,0 +1,15 @@ +[gene-expression] +reference,/path/to/cellranger/prebuilt/human/refdata-gex-GRCh38-2020-A/ +cmo-set,/path/to/working-directory/demultiplexing/5p_hashing_demux_cmo-set.csv + +[libraries] +fastq_id,fastqs,feature_types +PBMC-ALL_60k_universal_HashAB1-4_BL_4tags_Rep1_gex,Path/to/GEX/FASTQs,Gene Expression +PBMC-ALL_60k_universal_HashAB1-4_BL_4tags_Rep1_ab,Path/to/AB/FASTQs,Multiplexing Capture + +[samples] +sample_id,cmo_ids +Donor1_healthy,Hash-tag1 +Donor2_healthy,Hash-tag2 +Donor3_ALLpatient,Hash-tag3 +Donor4_ALLpatient,Hash-tag4