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自制的网站定向诱变整个质粒
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Biologia
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JoVE Journal Biologia
Homemade Site Directed Mutagenesis of Whole Plasmids
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自制的网站定向诱变整个质粒

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

网站定向诱变整个质粒是一种简单的方法来创建原始质粒略有不同的变化。在这里，我们展示了一个简单而成本效益的方法，引入碱基质粒使用的标准试剂。

Tags

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

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