Random insertion of T7 promoter to drive gene expression with T7 RNA polymerase

• Insert T7 promoter randomly into any DNA sequence in vitro
• Isolate positive clones with the selectable kanamycin marker
• Express resulting products in vivo or in vitro
• Minimize insertion bias with the hyperactive Tn5 system, known for highest level of randomness

Applications

• Random insertion of a single T7 RNA Polymerase promoter to synthesize RNA from any region of cloned DNA.

The EZ-Tn5™ Promoter Insertion Kit* provides an easy and reliable method to randomly insert a transposon containing a phage T7 RNA polymerase promoter into any DNA molecule in vitro. The transposon end has no termination sequences, so RNA can be produced from chosen insertion clones by in vitro transcription from the T7 RNA polymerase promoter using, for example, any of Epicentre\'s T7 RNA Polymerase Transcription Kits. RNA can also be generated for in vivo expression studies in cells having an inducible T7 RNA polymerase gene.

Figure 1 (click to enlarge). RNA transcripts can be produced from insertion clones in vitro or in vivo using the EZ-Tn5™ Promoter Insertion Kit.

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

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