Insert a selectable marker for sequencing and functional analysis of cloned DNA.

• Insert a kanamycin, tetracycline, or DHFR selectable marker into any DNA sequence in vitro
• Completely sequence cDNA or genomic clones in plasmid, cosmid, fosmid, or BAC vectors without primer walking
• Speed functional analysis without subcloning - create libraries of random mutants from purified DNA
• Minimize insertion bias with the hyperactive modified Tn5 system, known for lowest insertion bias

Simpify mutagenesis of cloned genes, and insert primer "landing pads" for easy sequencing of large genes with in vitro transposon tools

EZ-Tn5™ Insertion Kits are designed to simplify and speed up complete sequencing of any cloned DNA >2 kb, without primer walking or subcloning. Perform a simple, one-step in vitro reaction for random insertion of a single EZ-Tn5 Transposon containing a selectable marker—either kanamycin resistance (KAN), tetracycline resistance (TET), or trimethoprim resistance encoded by the dihydrofolate reductase gene (DHFR)—into the clone. Then, transform E. coli cells with an aliquot of the reaction and select for the marker encoded by the EZ-Tn5 Transposon. Use the primers provided in the kits to sequence insertion clones bidirectionally from primer binding sites at the transponson ends.

A single reaction generates up to 10⁶ insertion clones - enough to sequence even the largest clone, while saving time usually spent subcloning and designing sequencing primers. Multiple DNA samples can be processed in parallel, making in vitro transposomics an effective component of high-throughput sequencing pipeline for metagenomics research.

Covered by issued and/or pending patents, exclusively licensed or assigned to Epicentre® (an Illumina® Company).