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Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins
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JoVE Journal Genetica
Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

DOI:
10.3791/55312-v

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08:14 min

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February 25, 2017

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,
1Department of Biological Chemistry and Molecular Pharmacology,Harvard Medical School

Capitoli

  • 00:05Titolo
  • 00:34Multiple-turnover Primer Synthesis Reaction by Manual Sampling
  • 02:12Multiple-turnover Primer Synthesis Reaction Using a Rapid-quench Instrument
  • 06:11Results: Kinetics of Lagging-strand DNA Synthesis Revealed
  • 07:43Conclusion

Summary

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We describe sensitive, gel-based discontinuous assays to examine the kinetics of lagging-strand initiation using the replication proteins of bacteriophage T7.

Tags

DNA SynthesisLagging-strandBacteriophage T7DNA ReplicationDNA PrimaseDNA PolymeraseOkazaki Fragment SynthesisRadioactive MaterialsFormamide Dye BufferGene Four ProteinGP Four Primase HelicaseMagnesium ChlorideDenaturing Gel ElectrophoresisRapid Quench Flow Instrument

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