Contents > Transposomics™ > EZ-Tn5™ <T7/KAN-2> Promoter Insertion Kit
EZ-Tn5™ <T7/KAN-2> Promoter Insertion Kit
Supplier: EPICENTRE® Biotechnologies
EZ-Tn5™ <T7/KAN-2> Promoter Insertion Kit
| Catalogue number | Pack size |
| EZI03T7 | 10 reactions |
EZ-Tn5™ Transposase
| Catalogue number | Pack size |
| TNP92110 | 10 reactions |
TransforMax™ EC100™ Electrocompetent E. coli*
| Catalogue number | Pack size |
| EC10005 | 5 X100µl |
| EC10010 | 10X100µl |
*TransforMax™ EC100™ Electrocompetent Cells are recommended for high-efficiency, non-size-biased cloning of DNA containing EZ-Tn5™ Transposon insertions.
For Research Use Only. EZ-Tn5™ products covered by issued and pending patents
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Last modified: 2008-07-15 14:57:15
Synthesise RNA from any region of your cloned DNA
The EZ-Tn5™ <T7/KAN-2> 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. A simple two hour in vitro reaction randomly inserts a single EZ-Tn5 <T7/KAN-2> Transposon into the plasmid, cosmid, fosmid or BAC clone. Then, following transformation into E. coli, transposon insertion clones are selected on kanamycin plates. The insertion site of individual kanamycin-resistant insertion clones can be mapped or bidirectionally sequenced using the two sequencing primers included in the kit. 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, AmpliScribe™ T7 High Yield Transcription Kit, AmpliCap™ T7 High Yield Message Maker Kit, DuraScribe® T7 Transcription Kit or T7 RNA Polymerase. RNA can also be generated for in vivo expression studies in cells having an inducible RNA polymerase gene.
 | Figure 1. RNA transcripts can be produced from insertion clones in vitro or in vivo using the EZ-Tn5™ <T7/KAN-2> Promoter Insertion Kit.. |
Last modified: 2008-05-14 16:46:46
Benefits
- Produce RNA in vitro from the T7 RNA polymerase promoter in each randomly inserted transposon
- Produce RNA in vivo, from the promoter, after transformation of chosen transposon insertion clones into E. coli harbouring an inducible T7 RNA polymerase gene [e.g. BL21(DE3)pLysS]
- Generate a population of >106 T7 promoter insertion clones - each with a single randomly inserted T7 promoter
- EZ-Tn5 <T7/KAN-2> Transposon insertions are highly random
Last modified: 2008-05-14 16:46:46
Protocols for: EZ-Tn5™ <T7/KAN-2> Promoters Insertion Kit
EZ-Tn5™ <T7/KAN-2> Promoters Insertion Kit Protocol
(catalogue number EZI03T7)
EZ-Tn5™ <R6Kγori/KAN-2> Transposon, KAN-2 FP-1 Forward Primer & R6KAN-2 RP-1 Reverse Primer Protocol
