293T transfection protocolPEI transfection protocolStable clone selection protocolStable transfection protocolLinear DNA transfectionLinearize plasmid before transfectionGeneration of a stable cell line - Lonza Knowledge Center
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Generation of a stable cell line: A positive control plasmid works, ... (Linearized) plasmid should always be checked on an agarose gel for integrity before ... Q: Generationofastablecellline:Apositivecontrolplasmidworks,butIcannotobtainstablecloneswithmyconstructofinterest.Whatcouldbethereason? A: Onereasoncouldbethatyourgeneproductistoxictothecells.WerecommendtestingtheconstructforexpressionofthegeneofinterestbytransientNucleofection™ofyourcells.Iftheproportionofexpressingcellsdropsbetween4and48hourspostNucleofection,thiscanbeanindicationofyourgeneproductbeingtoxic.AdifferentreasoncouldbethattheDNAisdegraded.(Linearized)plasmidshouldalwaysbecheckedonanagarosegelforintegritybeforeNucleofection. RelatedURLs: RelatedLiterature: Lonza_BenchGuides_Guideline_for_Generation_of_Stable_Cell_Lines__Technical_Reference_Guide.pdf Categories: Transfection ResearchAreas: Uncategorized Youmayalsowanttoknow: CanIuseNucleofection™tocreateastableknockdowninacancercellline? HowmuchDNAshouldIuseinmyNucleofection™Reactiontoobtainstableclones? Howlongdoesittaketoobtainstabletransfectants? FollowingNucleofection™,Icannotobtainstableclonesfromsinglecells,whatcouldbethereason? HowmanystablecloneswillIgetfromonetransfection? WhatselectionmarkerscanIuseforstabletransfections? HowlongistheMycoAlert™Assaysignalstable? WhyistheNucleofector™Technologyidealforproteinproductionfromstableclones? WhyistheNucleofector™Technologyidealforprimarycellsanddifficult-to-transfectcelllines? IsyourpmaxGFPcontrolvectorsuppliedwiththeNucleofector™Kitssuitableforstableexpression? SeeMore
延伸文章資訊
- 1Linearization of plasmid DNA before stable transfection - blunt ...
I want to linearize a plasmid and just realized that there is no blunt end restriction site avail...
- 2Stable Transfection - Mirus Bio
When generating a stable cell line, the transfected plasmid undergoes recombination during chromo...
- 3Linearized or unlinearized plasmid for stable transfection?
If I didn't cut the plasmid to do stable transfection, ... a case who used linearised DNA and she...
- 4Guidelines for Plasmid DNA Transfection - TW
Stable transfections are more efficient when using linear DNA due to its optimal integration into...
- 5Stable transfection of MSCs by electroporation - Nature
After electroporation of 106 cells with. 10 mg of the linearized plasmid DNA, hMSCs with stable D...
