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Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
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Biochemistry
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JoVE Journal Biochemistry
Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

DOI:
10.3791/58996-v

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07:04 min

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February 05, 2019

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, ,
1Department of Biochemistry and Molecular Biology,University of Southern Denmark

Chapters

  • 00:04Title
  • 00:27PCR Amplification of Wild Type DNA for Cloning
  • 01:54Site-directed Mutagenesis – 2-step PCR
  • 03:01Site-directed Mutagenesis – 3-step PCR
  • 04:10Results: Investigating RNA Interactions using 2-step and 3-step Site-directed Mutagenesis Based Approach
  • 06:29Conclusion

Summary

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Site-directed mutagenesis is a technique used to introduce specific mutations in deoxyribonucleic acid (DNA). This protocol describes how to do site-directed mutagenesis with a 2-step and 3-step polymerase chain reaction (PCR) based approach, which is applicable to any DNA fragment of interest.

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Keywords: Site-directed MutagenesisRNA InteractionsEscherichia ColiPCRDNA TemplatePrimer PairsAgarose Gel ElectrophoresisDNA PurificationSpectrophotometry2-step PCR3-step PCRRNA-RNA InteractionsRNA-protein Interactions

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