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Chapters
- 00:04Title
- 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
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
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
