Journal
/
Génétique
/
Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1
Journal JoVE
Génétique
This content is Free Access.
Journal JoVE Génétique
Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1

Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1

DOI:
10.3791/60045-v

•

11:27 min

•

September 18, 2019

•

, , , , , , , ,
1Laboratory of DNA Replication and Recombination, Biological and Environmental Sciences and Engineering Division (BESE),King Abdullah University of Science and Technology (KAUST)

Chapitres

  • 00:04Titre
  • 01:22Flow Cell Preparation
  • 03:01Single-Molecule Förster Resonance Energy Transfer (smFRET) Experiments
  • 06:25Sm-FRET Data Analysis
  • 07:10Two-Color Alternating Excitation (ALEX) FRET Experiment and Analysis
  • 08:24Results: Representative Real-Time HJ Resolution Investigation
  • 10:13Conclusion

Summary

Traduction automatique

Traduction automatique

Presented here is a protocol for performing single-molecule Förster resonance energy transfer to study HJ resolution. Two-color alternating excitation is used for determining the dissociation constants. Single-color time lapse smFRET is then applied in real-time cleavage assays to obtain the dwell time distribution prior to HJ resolution.

Tags

Single-moleculeFörster Resonance Energy Transfer (FRET)Holliday JunctionGEN1Real-time InvestigationEnzymatic SystemsSingle-molecule MethodsBiomolecular InteractionsOptical And Force MeasurementsSingle-channel Flow CellQuartz SlideDouble Adhesive SheetPolytetrafluoroethylene TweezersPolyethylene TubingEpoxy GlueAvidinPBS

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

check_url/fr/60045?article_type=v

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
check_url/fr/60045?article_type=v

Detection of microRNA Expression in Peritoneal Membrane of Rats Using Quantitative Real-time PCR
check_url/fr/60045?article_type=v

Single Molecule Analysis of Laser Localized Psoralen Adducts
check_url/fr/60045?article_type=v

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
check_url/fr/60045?article_type=v

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
check_url/fr/60045?article_type=v

Single Molecule Fluorescence In Situ Hybridization (smFISH) Analysis in Budding Yeast Vegetative Growth and Meiosis
check_url/fr/60045?article_type=v

Identifying Mutations by High Resolution Melting in a TILLING Population of Rice
check_url/fr/60045?article_type=v

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii
check_url/fr/60045?article_type=v

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells
check_url/fr/60045?article_type=v

Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells

Read Article