Method for efficient soluble expression and purification of recombinant hyperactive Tn5 transposase
Tongtong Xu1, Minqin Xiao1, Lin Yu1
1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center of Biotherapy, 610041, Chengdu, China.
Abstract:
Efficient preparation of libraries is the key step of next-generation sequencing (NGS) methods. Tn5 transposase enables simple, robust and highly efficient tagmentation-based library construction. Here, we report a simple and reliable expression and purification strategy based on fusing Tn5 to the small B1 immunoglobulin binding domain of Streptococcal protein G (GB1) and high affinity 10× His tag. The purified recombinant Tn5 showed high DNA tagmentation activity and ultra-low nucleic acid contamination. This method greatly cuts the costs of Tn5-based NGS library construction and is beneficial to the development of new NGS methods.


