Insert an R6Kγ origin and kanamycin resistance for mutagenesis and rescue of flanking DNA.

• Insert R6K origin and kanamycin resistance marker randomly into any DNA sequence in vitro
• Characterize non-E. coli plasmids or circular genomes
• Simplify Sanger sequencing of large DNA inserts
• Minimize insertion bias with the hyperactive modified Tn5 system, known for lowest insertion bias

Characterize circular genomes and plasmids with in vitro transposon tools

The EZ-Tn5™ Insertion Kit facilitates the sequencing and genetic analysis of plasmids or any other circularized DNA that would not otherwise replicate in E. coli. The kit is based upon the EZ-Tn5™ Transposon which carries the E. coli R6Kγ conditional origin of replication (R6Kγori) and a kanamycin resistance marker. Use a simple one-step, 2-hour in vitro reaction to randomly insert the transposon into the target DNA. Then transform an aliquot of the reaction into an E. coli host expressing the pir gene product, which is required for replication from the R6Kγori. Select insertion clones on kanamycin plates and sequence them bidirectionally using the provided sequencing primers that are homologous to the transposon ends. Clones can be maintained in Epicentre's TransforMax™ EC100D™ pir+ or TransforMax™ EC100D™ pir-116 strains.

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

Randomly insert an R6Kγ origin and kanamycin resistance marker (the provided transposon sequence) into any purified plasmid or genomic DNA to create a library of easily-to-sequence mutants for a host of applications, including rescue of flanking DNA.