Application of Tn5 Transposase: The Fields, Materials, and Methods

      As an enzyme that has several applications in the field of biotech, Tn5 transposase, a bacterial transposase working in a “cut-and- paste” mode with amplification, can insert specific DNA sequences into a variety of substrates, including plasmids, bacterial chromosomes, and artificial DNA constructs. The following content will introduce the usage method of Tn5 transposase and the application of Tn5 transposase into genomic research and pharmaceuticals. 

      To get ready for any further application, Tn5 tagmentation is a technique used in DNA sequencing to prepare DNA fragments for library construction. The Tn5 transposase is an enzyme that can simultaneously cut and ligate DNA, allowing for the creation of small DNA fragments with adaptors attached to the ends. 

Buffer Construction

1.1 Dissolve your reference primer A, B, and ME with Annealing Buffer 

      Tagmentation-based library preparation of full-length RNA sequencing using Tn5 needs to be performed in 10mM Tris-HCl pH 7.5, 10mM MgCl2. And 25% dimethylformamide using 100-200pg cDNA. 

1.2 Mix Reaction 1 (Primer A solution) and Reaction 2 (Primer B solution) by vortexing, and collect the solution.

      The fragmentation is usually achieved at a 20-40 ng/ul concentration, incubated at 55 Celsius degree for 3 minutes in a preheated thermocycler. PCR amplification needs to be prepared at the same time. The residual dNTPs, oligonucleotides, and polymerase were cleared by bead purification [4].

Application to Genomic Research

      When talking about genomic research, there are several terms that we need to exemplify, such as NGS (next generation sequencing technology); ATAC-seq (assay for transposase accessible chromatin); and LIANTI “linear amplification via transposon insertion”. With the aid of construction of large libraries of diverse DNA sequences, these libraries can then be expressed and screened for compounds with desired chemical properties, such as a particular accession of protein. 

      Tn5 has its fundamental role when it comes to accessiona and amplification of DNA. It helps prepare the process of tagmentation, helps create the ATAC-seq for transcription or translocation.

      The transposase can also be massively applied for creation of artificial DNA constructs for use in nanotechnology and materials science. It helps modify a desired size, quantity, and traits of an artificial DNA by inserting specific sequences into a scaffold material. The nanostructures created in this way can have potential utility in the field of electronics. photonics, and biomedical engineering. This method goes along with Tn5 tagmentation application of genome-wide profiling of protein binding and genomic marks [2]. Protein contains enormous information. Application of Tn5 allows us to access and transfer this information and to get it used in transposase reactions with PCR primers or in ChIP-seq interactions (between protein and chromatin).

      Tn5 transposase can efficiently form complexes in vitro with the assistance of enzymatic activities. It’s usually not affected by bidding sites bias. In this way, while creating hybrid molecules, the transposases are easily guided to specific regions of the genome. This mechanism helps develop various methods of DNA modification with lower costs, turnover, and more applicable feedback.

Application to Pharmaceutical

      Tn5 can as well be used for facilitation of screening and lead optimization in drug industry with its trait of being used to create large libraries of compounds that were mentioned previously. Tn5 transposase’s trait of “cut-and-paste” helps to isolate the fragment and start the modification without any other costs. It also goes along with another application of Tn5 transposase, which is the creation of GMOs. With its fundamental function of inserting and amplifying DNA sequences, it can produce therapeutic proteins in large quantities and be affordable and accessible at the same time. More importantly, Tn5 transposases can be used in the development of targeted therapies for genetic diseases in this way. Genetic mutations that cause disease might be able to be fixed by applying the transposases ’traits of insertion, modification, and amplification. Severe genetic disease such as cystic fibrosis and sickle cell anemia can be detected or even fixed in the early stage of prenatal.

      With a wide range of applications in molecular biology and biotechnology, Tn5 has overall proven to be a versatile and accessible tool in the field. Its leading advantages in genome sequencing, gene expression analysis, mutagenesis, and genome engineering makes it a trendy choice for research and production use. To find out more about qualified source of Tn5 transposase, please visit www.betalifesci.com.