- Convenient,pre-processedvectorseliminatetheneedfordigestion,gelpurification,anddephosphorylation.
- TranscriptionterminatorsflankingcloningsitestABIlizeotherwisetoxicinserts.
- Choiceofcopynumberandantibioticresistance.
- OptimizedforusewithE.cloni®10Gelectrocompetentandchemicallycompetentcells(soldseparately)
Clonequicklyandfacilitatedownstreamapplications
Lucigen’suniquecloningkitswithpre-processedvectorsandhighlycompetentcellseliminatehoursoftediouspreparationandenableefficientcloning.Ligationcanbecompletedinaslittleas30minuteswithnoclean-upneeded;onlyabriefheatdenaturationisnecessarybeforetransformation.Downstreammanipulation(e.g,mutagenesis,subsequentinsertaddition)isfacilitatedbytheminimalvectorsize(1.7-2.0kb).
StabilizedInsertsandGap-FreeCloning
Allcloningkitsincludethetranscription-andtranslation-freepSMART®vectors(Figure1),whicheliminatemanyoftheproblemsassociatedwithcloningrecalcitrantDNAinconventionalplasmidvectors.Conventionalcloningvectorscontainpromoters(e.g.,lacZpromoter)thatconstitutivelytranscribetheinsertsequence.Thistranscriptionandcoupledtranslationcandestabilizeinsertsthatcontaincodingregions,strongpromoters,shortrepeats,orincompatIBLesecondarystructures.ThecloningsiteofpSMARTvectorsdoesnotincludealaczsequence.Inaddition,strongtranscriptionterminatorsflankthecloningsitetoblockspurioustranscriptionfromthevectorandinsert-driventranscriptionintothevector.LowcopynumberversionsofpSMARTfurtherstabilizesequencesthataredifficulttomaintainintypicalvectors.
NoBackgroundProblems
ThepSMARTvectorsaresuppliedpre-digested,withblunt,dephosphorylatedends,andarequalifiedtoproduce99.5%recombinantclonesintypicalexperiments.Theultra-lowbackgroundofemptyvector(lessthan0.5%)eliminatestheneedtoscreenforrecombinants,removestheuncertaintyoffalsenegatives(lightbluepUCcolonies)andfalsepositives(whitecoloniesthatlackinserts),andenableslibraryconstructionfromnanogramamountsofDNA.
Figure1.Highcopy(HC)andlowcopy(LC)versionsofthepSMARTtranscription-freecloningvectors.
ConvenientSuccess
EfficientlyobtainyourcloneorcreateyourgenomicorCDNAlibrary—theCloneSmartCloningKitseliminatetediouspreparationofvectorsandcompetentcells,aswellastime-consumingQCtestingexperiments.Kitsincludeoptimizedreagents,ligation-readyvector(nopost-ligationcleanupsteprequired),detailedinstructions,andtrouble-shootingguidestosimplifycloningandsequencing.HighlyefficientE.cloni®10Gelectrocompetentcells(upto>4×1010cfu/µg)orchemicallycompetentcells(>1×109cfu/µg)areavailableforsaleseparately.
