{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"submitter":["Simin Wan"],"organism":["Mus musculus"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16000"],"description":["To analyze TCR diversity, CD8+ T cells specific to the OVA (257-264) peptide-H-2Kb tetramers were isolated from Cd4-Cre and Ch25hfl/flCd4-Cre mice following VSV-OVA infection. The KLRG1+CD127– effector cell population was subsequently sorted, and total RNA was extracted from the cell samples using TRIzol reagent."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Growth Protocol - Mice were housed under a 12-hour light-dark cycle, at temperatures of 18–22°C, and at a humidity of 30–70%.","Library Construction - Libraries were constructed from total RNA using a 5' RACE approach for reverse transcription, followed by TCR-specific PCR amplification. The PCR products were purified, end-repaired, and ligated to sequencing adapters. The final libraries were amplified with P5 and P7 primers, purified, and quality was assessed using a Qsep100 and Qubit 3.0 Fluorometer.","Sample Collection - KLRG1+CD127- effector cells were sorted from OVA257-264-H-2Kb tetramer-binding CD8+ T cells isolated from Ch25hfl/fl and Ch25hfl/flCd4-Cre mice after infection with VSV-OVA. Cell sorting was performed on a BD FACSAria IIu flow cytometer.","Sample Treatment - Mice were infected intravenously with 1 × 10^6 plaque-forming units (PFU) of recombinant VSV-OVA.","Nucleic Acid Extraction - Total RNA was extracted from the sorted KLRG1+CD127- effector cell samples using TRIzol reagent.","Sequencing - Libraries were multiplexed and sequenced on an Illumina MiSeq or NovaSeq instrument using a 2x250 or 2x300 paired-end configuration. Image analysis and base calling were conducted by the MiSeq Control Software (MCS) or NovaSeq Control Software (NCS)."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"data_protocol":["Sequence Alignment - Raw FASTQ files were quality-trimmed using Trimmomatic (v0.36). Paired-end reads were merged using FLASH (v2.2.00). Merged sequences were aligned to the IMGT reference database using BLAST for germline V(D)J gene assignment and CDR region annotation.","Data Transformation - TCR clonotype frequencies were normalized to counts per million (CPM). Diversity indices (Observed Clonotypes, Chao1, Shannon, Simpson) were calculated based on these normalized frequencies."],"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"instrument_platform":["Illumina MiSeq"],"study_type":["RNA-seq of coding RNA"],"species":["Mus musculus"],"pubmed_authors":["Jianhua Li","Simin Wan"],"additional_accession":[]},"is_claimable":false,"name":"TCRb-seq of mouse CD8+ T cells isolated from Cd4-Cre and Ch25hfl/flCd4-Cre mice following VSV-OVA infection","description":"To analyze TCR diversity, CD8+ T cells specific to the OVA (257-264) peptide-H-2Kb tetramers were isolated from Cd4-Cre and Ch25hfl/flCd4-Cre mice following VSV-OVA infection. The KLRG1+CD127– effector cell population was subsequently sorted, and total RNA was extracted from the cell samples using TRIzol reagent.","dates":{"release":"2026-04-01T00:00:00Z","modification":"2026-04-02T01:05:02.573Z","creation":"2025-11-06T10:59:31.821Z"},"accession":"E-MTAB-16000","cross_references":{"ENA":["ERP183805"],"EFO":["EFO_0002944","EFO_0004170","EFO_0003789","EFO_0004917","EFO_0005518","EFO_0003816","EFO_0003738","EFO_0004184","EFO_0003969"]}}