One of the major issues in the production of home-brew Tn5 transposes is the availability of lab equipment such as HPLC facilities. Although some methods for producing home-brew Tn5 without the need of such facility have been published, it was reported to be quite difficult to replicate. We have also encountered difficulties while attempting to follow these exact methods. As such, changes to this method have been made to facilitate the process. We have integrated a large-scale production of pure Tn5 into our routine, which is then used for the assembly of Tn5 transposomes. This presentation provides a description of a novel Tn5 based NGS library construction method using a unique Tn5 transposome assembling strategy. We believe it will improve single cell based NGS applications such as single cell Assay for Transposase Accessible Chromatin Sequencing (ATAC-seq) and Whole Genome Bisulfite Sequencing (WGBS).
KEYWORDS: Tn5 transposase; Single cell barcode; ATAC-seq; Next Generation Sequencing (NGS)