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Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA
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생물학
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JoVE 신문 생물학
Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

DOI:
10.3791/56551-v

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12:35 min

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November 14, 2017

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, ,
1Department for Medical Biochemistry and Cell Biology,University of Gothenburg, 2Department of Biology and Biological Engineering,Chalmers University of Technology

Chapters

  • 00:05Title
  • 00:55HincII Treatment and Alkaline Hydrolysis
  • 04:115′-End Phosphorylation
  • 05:21Single Strand DNA Ligation
  • 06:35Second Strand Synthesis
  • 08:23PCR Amplification, Library Analysis and Pooling
  • 10:10Results: Simultaneous Quantitation and Genome-wide Mapping of Ribonucleotides
  • 11:39Conclusion

Summary

자동 번역

자동 번역

Here we describe a method amenable to simultaneously quantitate and genome-wide map ribonucleotides in highly intact DNA at single-nucleotide resolution, combining enzymatic cleavage of genomic DNA with its alkaline hydrolysis and subsequent 5´-end sequencing.

Tags

Mitochondrial DNARibonucleotidesDNA IntegrityDNA ReplicationDNA RepairHeLa CellsHincIIParamagnetic Beads70% EthanolPotassium HydroxidePotassium ChlorideSodium Acetate

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