- Createcustomtransposomesforinvitrotransposomics
- IntegrateselectableMarkers,promotersorreplicationoriginsintoDNAinvitrowithEZ-Tn5insertionkits
Applications
- InvitroinsertionofanEZ-Tn5TransposonintoDNAclonedinvectors,suchasplasmids,fosmids,cosmids,orBACsaswellasinvitroinsertionintolinearDNA.*
- PreparationofEZ-Tn5Transposomesforinvivotranspositionfollowingelectroporationintolivingcells.**
EZ-Tn5™TransposaseisahyperactiveformofTn5transposase.1Thehighlypurified,single-subunitenzymecanbeusedtorandomlyinsert(transposeor"hop")anyEZ-Tn5TransposonintoanytargetDNAinvitrowithanefficiencyupto>106insertionclonesperstandardreaction.WhenincubatedwithanEZ-Tn5TransposonintheabsenceofMg2+,astableEZ-Tn5Transposome™complexisformed.TheTransposomeissostablethatitcanbeelectroporatedintolivingcells.Onceinthecell,theTransposomeisactivatedbyintracellularMg2+andtheEZ-Tn5Transposoncomponentisrandomlyinsertedintothehost'sgenomicDNA.Thethree-dimensionalstructureofaTransposomecomplexhasbeenelucidated.2
AtypicalEZ-Tn5transpositionreactionrequiresfourcomponents:(1)theEZ-Tn5Transposase;(2)anEZ-Tn5Transposon;(3)atargetDNA;and(4)thepresenceofMg2+.ThehighlyrandominsertionofanEZ-Tn5TransposonintothetargetDNA3proceedsbyacut-and-pastemechanism,1catalyzedbytheEZ-Tn5Transposase,andresultsina9-bpduplicationoftargetDNAsequenceimmediatelyadjacenttobothendsoftheTransposon.
Benefits
- SpecificallyanduniquelyrecognizestheOuterEndsequencesofnaturallyoccurringTn5andmini-Tn5transposonsandthehyperactiveMosaicEndsofEZ-Tn5Transposons.
References
- Goryshin,I.Y.andReznikoff,W.S.(1998)J.Biol.Chem.273,7367.
- Davies,D.R.etal.(2000)Science289,77.
- Shevchenko,Y.etal.(2002)NucleicAcidsRes.30,2469.
*Coveredbyissuedand/orpendingpatents,exclusivelylicensedorassignedtoEpicentre®(anIllumina®Company).
**TheuseofTransposome™complexesforinvivoinsertionofatransposon,including,butnotlimited,toHyperMu™andEZ-Tn5™Transposome™complexes,iscoveredbyU.S.PatentNo.6,159,736andrelatedpatentsandpatentapplications,exclusivelylicensedtoEpicentre®(anIllumina®Company).
ORDERINFORMATION
10unitsoftransposaseisenoughenzymefor10standardreactions.
Pleasenote:IlluminarequiresthataTransposome™LicenseAcknowledgementissignedpriortoprocessingordersforEZ-Tn5™Transposase.Pleasecompletethisformbeforesubmittingyouronlineorderforthisitem.
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