Bettertransformationefficiencieswithrestrictionsysteminhibition
IncreasetheinsertionefficiencyofEZ-Tn5™Transposome™complexes.- EnhancethetransformationefficiencyofunmodifiedDNAbyblockingtypeIR-Msystems,whicharewidespreadinEubacteriaandArchaebacteria.
Applications
- IncreasethetransformationefficiencyofunmodifiedDNA2byblockingtypeIR-Msystems,whicharewidespreadinEubacteriaandArchaebacteria.3
- IncreasetheinsertionefficiencyofEZ-Tn5™Transposome™complexes.2
DNAtransformationcanbedifficulttoachieveinmanybacterialstrainsduetothepresenceofoneormorerestrictionandmodification(R-M)systemsthatcleaveunmodifiedDNAthatisrecognizedas"foreign."TypeOne™RestrictionInhibitor*providesapowerfulmethodforincreasingtransformationefficienciesinbacterialstrainswithtypeIR-Msystems.Developedasauniquepreparationofaphageproteincalledocr,1TypeOneInhibitorisreADIlyelectroporatedintocellsalongwithtransformingDNA.Invivo,theproteinactsasamoleculardecoythatblockstheDNA-bindingsiteoftypeIR-MenzymesandinhibitsDNAcleavage.
Benefits
- Easytouse:SimplyaddTypeOneRestrictionInhibitortounmodifiedDNAoraTransposomeandelectroporateusingstandardmethods.
- BlocksallknownfamiliesoftypeIR-Menzymes1andcanthereforebeusedtoincreasetransformationefficienciesinavarietyofbacterialcells.
Table1.EffectofTypeOne™RestrictionInhibitorontransformationefficiencies.
Strain(TypeIR-Msystem) | TypeOne™ Inhibitor | TypeofDNAorTransposome™ | Recombinants perµgDNA |
| S.typhimuriumLT2(StyLTIII) | pUC19(100pg) | 3.0×106 | |
| S.typhimuriumLT2(StyLTIII) | X | pUC19(100pg) | 3.0×108 |
| E.coliMG1655(EcoK1) | 48kbfosmid(50ng) | 3.0×103 | |
| E.coliMG1655(EcoK1) | X | 48kbfosmid(50ng) | 1.4×106 |
| S.typhimuriumLT2(StyLTIII) | EZ-Tn5™<R6Kγori/KAN-2>TnpTransposome™(1µl) | 1.3×104 | |
| S.typhimuriumLT2(StyLTIII) | X | EZ-Tn5™<R6Kγori/KAN-2>TnpTransposome™(1µl) | 1.0×106 |
References
- Walkinshaw,M.D.etal.(2002)Molec.Cell9,187.
- Hoffman,L.etal.(2002)EpicentreForum9(2),8.
- Murray,N.E.etal.(2000)Microbial.Molec.Biol.Rev.64,412.
*Coveredbyissuedand/orpendingpatents,exclusivelylicensedorassignedtoEpicentre®(anIllumina®Company).
