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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
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Biologia
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JoVE Journal Biologia
Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

DOI:
10.3791/58020-v

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09:04 min

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July 26, 2018

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, ,
1Genome Integrity and Structural Biology Laboratory,National Institute of Environmental Health Sciences, NIH, DHHS

Capitoli

  • 00:04Titolo
  • 00:49Alkaline Hydrolysis of Genomic DNA
  • 02:36Strand specific Southern Blot: Transfer
  • 04:37Strand-specific Southern Blot: Hybridization
  • 06:48Results: Assessment of Ribonucleotide Incorporation into DNA
  • 08:08Conclusion

Summary

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Traduzione automatica

Ribonucleotides are among the most abundant non-canonical nucleotides incorporated into the genome during eukaryotic nuclear DNA replication. If not properly removed, ribonucleotides can cause DNA damage and mutagenesis. Here, we present two experimental approaches that are used to assess the abundance of ribonucleotide incorporation into DNA and its mutagenic effects.

Tags

Keywords: Ribonucleotide IncorporationStrand-specific DetectionYeast GenomeRibonucleotide-induced MutagenesisAlkaline Gel ElectrophoresisSouthern BlottingGenomic DNAPotassium HydroxideAlkaline DNA Loading BufferAgarose GelSodium HydroxideEDTADNA Size MarkerNeutralization BufferSYBR Gold StainUV TransilluminatorAlkaline Transfer BufferNylon MembraneCapillary Action

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