Tnp Transposome™ Kit
HyperMu™ <R6K gamma ori/KAN-1> Tnp Transposome™ Kit
- Rescue cloning of transposon-mutagenized microbial genes.
- Rescue of plasmids from non-E. coli bacteria.
The HyperMu™ <R6Kγori/KAN-1>Tnp Transposome™ is the stable complex between HyperMu Transposase and a HyperMu Transposon containing an R6Kγ E. coli conditional origin of replication and a kanamycin selectable marker. The Transposome can be electroporated into living E. coli cells, where it is activated by intracellular Mg2+, and the HyperMu Transposon is randomly inserted into the host's genomic DNA.* Transposon insertion clones are selected on kanamycin plates or by screening for a desired gene "knockout" or loss of gene function.
The transposon insertion site can be sequenced directly using the provided primers that are homologous to the ends of the inserted HyperMu Transposon. Alternatively, the insertion site and its flanking DNA can be "rescued" as a plasmid and then sequenced.
Note: EPICENTRE recommends that researchers consider using an EZ-Tn5™ Transposome Kit in lieu of or in addition to a HyperMu Transposome Kit. An EZ-Tn5 Transposome typically generates 10-100 times more in vivo insertions than a HyperMu Transposome.
- Easily recover and propagate plasmids from diverse bacterial genera that will not replicate normally in E. coli.
- Simple rescue cloning process of mutagenized genes speeds up structure/function studies and sequencing.
- Random insertion of transposon DNA assures excellent coverage of entire bacterial chromosome.
- Rescue clones can be sequenced bidirectionally using the provided primers that are homologous to the ends of the transposon.
|Figure 1. The process for rescue cloning transposon insertion sites in genomic DNA using the HyperMu™ <R6Kγori/KAN-1>Tnp Transposome™ and TransforMax™ EC100D™ pir+ or TransforMax™ EC100D™ pir-116 Electrocompetent E. coli.|
*The use of Transposome™ complexes for in vivo insertion of a transposon, including, but not limited, to HyperMu™ and EZ-Tn5™ Transposome™ complexes, is covered by U.S. Patent No. 6,159,736 and related patents and patent applications, exclusively licensed to EPICENTRE.
|HyperMu™ <R6Kγori/KAN-1>Tnp Transposome™ Kit|
|Includes unlabeled forward and reverse sequencing primers.|