{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Hu H"],"funding":["Norwegian Research Council","Swiss National Science Foundation","European Union","Trond Mohn stiftelse","Ministry of Health and Care Services","SNSF Bridge"],"pagination":["pgaf006"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11759286"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["4(1)"],"pubmed_abstract":["Here, we present Link-Seq, a highly efficient droplet microfluidic method for combined sequencing of antibody-encoding genes and the transcriptome of individual B cells at large scale. The method is based on 3' barcoding of the transcriptome and subsequent single-molecule PCR in droplets, which freely shift the barcode along specific gene regions, such as the antibody heavy- and light-chain genes. Using the immune repertoire of COVID-19 patients and healthy donors as a model system, we obtain up to 91.7% correctly paired immunoglobulin heavy and light chains. Furthermore, we map the V(D)J usage and obtain sensitivities comparable with the current gold-standard 10× Genomics commercial systems while offering full flexibility in experimental setup and significant cost savings. A further uniqu"],"journal":["PNAS nexus"],"pubmed_title":["Targeted barcoding of variable antibody domains and individual transcriptomes of the human B-cell repertoire using Link-Seq."],"pmcid":["PMC11759286"],"funding_grant_id":["H2020 101037867 Vaccelerate","H2020 874866 INCENTIVE","F-11628","218721","40B2-0_218721","TMS2020TMT05","284930","EU IMI 2 101007799 Inno4Vac"],"pubmed_authors":["Brokstad KA","Hu H","Kolmar L","Ma X","Zhou F","Girardot C","Cox RJ","Benes V","Merten CA"],"additional_accession":[]},"is_claimable":false,"name":"Targeted barcoding of variable antibody domains and individual transcriptomes of the human B-cell repertoire using Link-Seq.","description":"Here, we present Link-Seq, a highly efficient droplet microfluidic method for combined sequencing of antibody-encoding genes and the transcriptome of individual B cells at large scale. The method is based on 3' barcoding of the transcriptome and subsequent single-molecule PCR in droplets, which freely shift the barcode along specific gene regions, such as the antibody heavy- and light-chain genes. Using the immune repertoire of COVID-19 patients and healthy donors as a model system, we obtain up to 91.7% correctly paired immunoglobulin heavy and light chains. Furthermore, we map the V(D)J usage and obtain sensitivities comparable with the current gold-standard 10× Genomics commercial systems while offering full flexibility in experimental setup and significant cost savings. A further uniqu","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Jan","modification":"2025-04-05T10:18:13.159Z","creation":"2025-04-05T10:18:13.159Z"},"accession":"S-EPMC11759286","cross_references":{"pubmed":["39867668"],"doi":["10.1093/pnasnexus/pgaf006"]}}