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Homemade Site Directed Mutagenesis of Whole Plasmids
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Biologia
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JoVE Journal Biologia
Homemade Site Directed Mutagenesis of Whole Plasmids

Homemade Site Directed Mutagenesis of Whole Plasmids

DOI:
10.3791/1135-v

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07:11 min

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May 11, 2009

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1Department of Biology,Johannes Gutenberg-University Mainz, Germany, 2Proteomics division, AlPlanta,Neustadt an der Weinstrasse, Germany

Capitoli

  • 00:05Titolo
  • 00:10Introduction
  • 00:47What We Need
  • 02:47Thermocycling Reaction
  • 03:29Thermocycling Conditions
  • 04:09Gel Check After Reaction
  • 04:28DpnI Digestion and Transformation
  • 04:58Transformation
  • 05:30Screening of Clones
  • 06:47Conclusion

Summary

Traduzione automatica

Traduzione automatica

Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. Here we demonstrate an easy and cost effective way to introduce base substitutions into a plasmid using standard reagents.

Tags

Site Directed MutagenesisWhole PlasmidsCloned Target GeneSubstitutionDeletionInsertionAmplifying PlasmidMutagenic PrimersBase SubstitutionsProtocolCost-effectiveCommercial KitsReaction Cost ReductionCritical StepsMutagenic Primer Design

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