ImprovedFosmidCloning
Lucigen’sCopyRight®v2.0FosmidCloningKitwiththepSMART®FOSvectorandReplicator™FOSstraincontainsthemostadvancedfosmidcloningdeveloped.TheCopyRightv2.0FosmidKityieldsmanymorerecombinantsfromlimitedstartingDNA(Figure1). ThevectorincorporatesCopyRighttranscriptionterminatorstostabilizeinserts,whichresultsincompletefosmidlibrarieswithoutmissingclones.False-negativesandfalse-positivesaredramaticallyreduced,andempty-vectorbackgroundisessentiallyeliminated(Figure2).ControllablegeneticelementsinthepSMARTFOSvector(Figure3)andReplicatorFOScellsallowamplificationofvectorcopynumberforhighplasmidyields(Figure4).
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Figure1.HigherrecombinantyieldswithalimitingamountofDNA. Lucigen’sCopyRightFosmidkitcomparedtoEpicentre’sCopyControlkit. |
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Figure2.ZerobackgroundwiththepSMARTFOSvector(left)comparedtotheCopyControl™pCC1FOS™vector(right)(Epicentre). |
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Figure3.CopyRightv2.0pSMARTFOSvector.sacB,sucrasegene;lacZ,lacZalphapeptidegene;Cm^r-chloramphenicolresistancegene;cosN-lamBDapackagingsignal;parA,B,C-partitiongenes;ori2,repE[notshownindrawing],IncC-single-copyoriginofreplication;oriV-inducibleoriginofreplication.Approximatepositionsofsequencingprimersandtranscriptionterminators(T)areindicated.ThelacZ/sacBstufferregioniscompletelyremovedduringvectorprocessing. |
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Figure4.CopynumberamplificationoffosmidclonesfromtheCopyRightv2.0FosmidKit(Replicatorcells)wasmoreconsistentandrobustthantheCopyControlFosmidkit(EPI300cells). |
ConvenientSuccess
Efficientlyobtainyourcloneorcreateyourlibrary—theCopyRightv2.0CloningKitseliminatetediousvectorandcompetentcellpreparation,aswellastime-consumingQCtestingexperiments. Kitsincludeoptimizedreagents,ligation-readyvector,optimizedcells,detailedinstructions,andtrouble-shootingguide.
ORDERINFORMATION
EachCopyRightv2.0FosmidCloningKitcontains: pre-cut,dephosphorylatedpSMARTFOSbluntvector,CloneSmartDNALigase,CloneDirect™10XLigationBuffer(includesATP),DNATerminatorEndRepairEnzymeMixandEndRepairBuffer,ReplicatorFOSstrain(glycerolstock),20%MaltoseSolution,1MMgSO4,SMBuffer,ArabinoseInductionSolution,SequencingPrimers,andcompleteprotocols.The20-reactionKitisprovidedastwo10-reactionKits.ebiomall.com
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般基克隆物体直接取基包括面几步骤
提取植物或者物基组DNA（试剂盒）————做PCR需要基扩增（顺利2搞定）——扩增基做酶切——基连接想插入载体面（顺利1-2）——鉴定克隆基
步骤都商品化试剂或者试剂盒 要思路清晰做起快 1-2周能完
没证实情况任何没依据推断都假想空想
现在小弟碰见一个难题，我想克隆一个基因，但是我在NCBI一检索，发现别人只发表了这个基因的DNA全长，但是，CDNA没有发表，也没有发表文章，想请问下，在不知道CDNA的情况下，请问我应该如何做这个基因的克隆？？？（全基因组也不知道,其他生物没有这种基因）
详见：http://zhidao.baidu.com/question/1831844001436785260.html?fr=iks&word=%B2%A1%B6%BE%B5%C4%B8%B4%D6%C6%D6%DC%C6%DA%B5%C4%B9%FD%B3%CC%3F&ie=gbk
