biostudies-literatureKuniyoshi YCore Research for Evolutional Science and Technology, Japan Science and Technology AgencyJapan Society for the Promotion of Sciencee0165473https://www.ebi.ac.uk/biostudies/studies/S-EPMC5082856biostudies-literatureUnknown11(10)Identification of immunoglobulin genes in hybridomas is essential for producing antibodies for research and clinical applications. A couple of methods such as RACE and degenerative PCR have been developed for determination of the Igh and Igl/Igk coding sequences (CDSs) but it has been difficult to process a number of hybridomas both with accuracy and rapidness. Here, we propose a new strategy for antibody sequence determination by mRNA-seq of hybridomas. We demonstrated that hybridomas highly expressed the Igh and Igl/Igk genes and that de novo transcriptome assembly using mRNA-seq data enabled identification of the CDS of both Igh and Igl/Igk accurately. Furthermore, we estimated that only 30,000 sequenced reads are required to identify immunoglobulin sequences from four different hybridoPloS oneIdentification of Immunoglobulin Gene Sequences from a Small Read Number of mRNA-Seq Using Hybridomas.PMC508285616K18479, 16H01577, 16H0155025116010, 2629006415K18457, 16H01219Iwasaki TKimura HFujita MMaehara KKuniyoshi YHarada AOhkawa YSato YHayashi MSemba YfalseIdentification of Immunoglobulin Gene Sequences from a Small Read Number of mRNA-Seq Using Hybridomas.Identification of immunoglobulin genes in hybridomas is essential for producing antibodies for research and clinical applications. A couple of methods such as RACE and degenerative PCR have been developed for determination of the Igh and Igl/Igk coding sequences (CDSs) but it has been difficult to process a number of hybridomas both with accuracy and rapidness. Here, we propose a new strategy for antibody sequence determination by mRNA-seq of hybridomas. We demonstrated that hybridomas highly expressed the Igh and Igl/Igk genes and that de novo transcriptome assembly using mRNA-seq data enabled identification of the CDS of both Igh and Igl/Igk accurately. Furthermore, we estimated that only 30,000 sequenced reads are required to identify immunoglobulin sequences from four different hybrido2016-01-01T00:00:00Z20162026-04-29T16:49:36.989Z2019-03-26T22:51:08ZS-EPMC50828562778822610.1371/journal.pone.0165473