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

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