- 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.
ORDERINFORMATION
CloneSmartBluntCloningKitscontain:VectorPremix,CloneSmartDNALigase,PositiveControlInsertDNA,SequencingPrimers,andacompleteprotocol.KitscanbeusedwithanyE.colichemicallycompetentorelectrocompetentcellsbuthavebeenoptimizedforusewithofE.cloni®ElectrocompetentCellsandChemicallyCompetentCells.ebiomall.com
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双缩脲试剂工作原理铜离强碱性条件与肽键发反应紫色络合物
（我做实验候斐林试剂先试管制加原糖溶液双缩脲试剂先往蛋白质加碱再加铜离所原双缩脲试剂硫酸铜浓度比氢氧化钠稀斐林试剂二者浓度相近原）
采用6孔板接毒后，铺上低溶解度琼脂糖，第二天就出来了这些病变，正常细胞没有出现这种现象。
按说空斑应该出现多个，但我的六孔板中一个空最多也就出现了2个空斑，也不像文献报道的那样规则，而且很大！
由于没有做梯度稀释，会不会是接毒量过大造成的啊？请大侠指点，万分感谢！
2) 克隆技术使用使倾向于量繁殖现种群利用价值体,按自规律促进整种群优胜劣汰.意义说,克隆技术干扰自进化程.
3) 克隆技术种昂贵技术,需要量金钱物专业士参与,失败率非高.莉277实验唯.虽现发展更先进技术,功率能达2-3%.
4) 转基物提高疾病传染风险.例,产药物牛奶牛染病毒,种病毒能通牛奶染病
5) 克隆技术应用于体导致代遗传性状工控制.克隆技术引起争论核能否允许发育初期类胚胎进行遗传操作.伦理家所能接受.
6) 克隆技术用创造超,或拥健壮体格却智力低.且,克隆技术能够类效运用,男性失遗传意义.
7) 克隆技术家庭关系带影响巨.由父亲DNA克隆孩看作父亲双胞胎兄弟,延迟几十已.难设想,发现自另外完全复制品,（或）受
