TMI-seq: Tn5 Transposase Mediated Production of Complex Libraries for Short Read Sequencing

Tech ID: 32761 / UC Case 2015-974-0


Although Next Generation Sequencing has vastly improved sequencing throughput while reducing sequencing costs, preparation of nucleic acid libraries for sequencing has become a bottleneck. In addition, it is difficult using short read next generation sequencing to assemble highly variable sequences that exceed 500 base pairs such as cDNAs derived from antibody heavy chain, antibody light chain, and T cell variable regions RNA.  

Technology Description

The technology, termed “TMI-seq” is a library preparation that combines molecular barcoding of individual molecules with “tagmentation” – a process by which the TN5 transposase both fragments and adds tags to DNA. In one use of the method, RNA is reverse transcribed to integrate a molecular barcode and a partial forward and reverse sequencing primer. It is then amplified by PCR, which integrates index sequences and full forward and reverse sequencing primers to yield a full length cDNA library. One aliquot of the library is tagmented with Tn5 and the forward sequencing primer, resulting in a library of barcoded overlapping fragments focused on the 5’ region of the target sequence. A second aliquot is tagmented with Tn5 and the reverse sequencing primer, resulting in a library of barcoded overlapping fragments focused on the 3’ region of the target sequence. 



  • RNA à cDNA library sequencing
  • Sequencing of T Cell receptors and antibodies



  • Fast prep and accurate assembly for short read sequencers like Illumina sequencers. 
  • High quality data from immune repertoire sequences
  • Accurate TCR and antibody repertoire sequences


Intellectual Property Information

Country Type Number Dated Case
United States Of America Issued Patent 11,319,576 05/03/2022 2015-974
United States Of America Issued Patent 10,280,449 05/07/2019 2015-974

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  • Cole, Charles
  • Vollmers, Christopher

Other Information


Illumina (R) sequencing, Short Read Sequencing, Transposase, Tn5, TMI-Seq, Antibody sequencing, Immune Repertoire Sequencing, cDNA libraries

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