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当前位置: 首页 > 产品中心 > CRISPR, Large or Difficult Fragment Cloning > Lucigen/CopyCutter™ EPI400™ Chemically Competent E.coli/C400CH10/10 x 50 µl
公司介绍
美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等
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Lucigen/CopyCutter™ EPI400™ Chemically Competent E.coli/C400CH10/10 x 50 µl

Lucigen/CopyCutter™ EPI400™ Chemically Competent E.coli/C400CH10/10 x 50 µl
  • Lucigen/CopyCutter™ EPI400™ Chemically Competent E.coli/C400CH10/10 x 50 µl
商品介绍

OptimizedforcloningtoxicorunstableDNA

  • Formerly from EpicentreStABIlizetoxicinsertsincommoncloningandexpressionvectors(pUCandpET-typevectors)
  • Cloneandmaintainchallengingsequencesatreducedplasmidcopynumber,theninducetohighcopynumberforDNArecovery
  • AvoidT1andT5phagecontaminationwithtonAmutation
  • Choosechemicallycompetentcellsforgeneralcloningorelectrocompetentcellsfordemandingapplicationssuchaslibrarygeneration

Applications

  • CloningofunstableDNAsequencesorthoseexpressingtoxicproteins.

CopyCutter™EPI400™E.coli*cellsweredevelopedtosignificantlylowerthecopynumberofawidevarietyofcommonvectorssothatyoucanmorereADIlycloneunstableDNAsequences.DNAthatisunstableathigh-copynumberoftencodesforaproteinthatinhibitscellgrowthorcontainsAT-andGC-richsequencesorsequenceswithstrongsecondarystructure(Fig.2).1

TheCopyCutterEPI400celllinewasderivedfromEpicentre'shigh-transformationefficiencyphageT1-resistantTransforMAX™EC100™-T1RE.colistrainbymanipulatingagenethatcontrolsthecopynumberofvectorscontainingColE1orpMB1originsofreplication(e.g.,pUC-andpET-typevectors).Thisconstitutivelyexpressedgene,pcnB(plasmidcopynumber),wasdeletedfromtheTransforMAXEC100strainandreplacedwithamodifiedpcnBgenethatislinkedtoaninducIBLepromoter,creatingtheCopyCutterEPI400strain.

ThecopynumberofColE1-typevectorsintheCopyCutterEPI400straincomparedtotheparentalTransforMAXEC100strainisapproximately4-to25-foldlower,dependingonthevector.Moreover,ashortincubationinthepresenceoftheCopyCutterInductionSolutioncanincreasethecopynumberofthevectortoimproveplasmidyields(Fig.3).

Figure1.Copy-numberofColE1-typeplasmidsisloweredupto25-foldinCopyCutter™EPI400™E.colicells.LanesC,TransforMax™EC100™cells;LanesUandI,uninducedorinducedCopyCutterEPI400cells.DNAextractsfromthesamenumberoflysedcells(basedonOD600)wereloadedperlane.

Figure2.DNAinsertsencodingtoxicgeneproductsweresuccessfullyclonedintohigh-copy-numbervectorsusingCopyCutter™EPI400™E.colicells.Aftersequencing,thefull-lengthacpPclonesinTransforMAX™EC100™cellswerefoundtocontainmultiplepointmutations.Figure3.UninducedCopyCutter™EPI400™E.colicellscontainingaregBclone(laneU)areinducedtohigher-copynumber(laneI)usingtheCopyCutterInductionSolution.CrudeextractsofplasmidDNAwerepreparedfromcellsgrowninselectivemediaandanalyzedbyagarosegelelectrophoresis.Approximatelythesamenumberoflysedcells(basedonA600)wereloadedperlane.

Benefits

  • Maintainclonesatlow-copynumber,theninducetohighercopynumberforimprovedplasmidyield.
  • Hightransformationefficiencywithclonesofallsizes.
  • tonAforresistancetobacteriophagesT1andT5.
  • lacZΔM15forblue/whitescreeningofrecombinants.
  • Restrictionminus[mcrA,Δ(mrr-hsdRMS-mcrBC)]genotypeenablesefficientcloningofmethylatedDNA.
  • Endonucleaseminus(endA1)toensurehighyieldsofDNA.
  • Recombinationminus(recA1)forgreaterstabilityoflargeclonedinserts.

Genotype

F-mcrAΔ(mrr-hsdRMS-mcrBC)Φ80dlacZΔM15ΔlacX74recA1endA1araD139Δ(ara,leu)7697galUgalKλ-rpsL(StrR)nupGtrfAtonApcnB4dhfr

CopyCutterEPI400ElectrocompetentE.coli

  • Transformationefficiencyof>1x1010cfu/µgofpUC19.

CopyCutterEPI400ChemicallyCompetentE.coli

  • Transformationefficiencyof>1x107cfu/µgofpUC19.

Reference

  1. Haskins,D.(2004)EpicentreForum11(5),6.

*Coveredbyissuedand/orpendingpatents.


ORDERINFORMATION

Tentubeseachcontaining50µLofcells(enoughcellsfor10transformations),CopyCutter™InductionSolutionandpUC19controlDNA.TheCopyCutter™InductionSolutionisprovidedata1,000Xconcentrationandisfiltersterilized.
品牌介绍

美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术,过硬的产品质量,良好的产品服务赢得了全球广大用户的信赖。


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美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术,过硬的产品质量,良好的产品服务赢得了全球广大用户的信赖。

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