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Genetics
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单分子Fürster共振能量传递方法,用于GEN1对霍利迪结分辨率进行实时调查
JoVE Journal
Genetics
This content is Free Access.
JoVE Journal
Genetics
Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1
Please note that all translations are automatically generated.
Click here for the English version.
单分子Fürster共振能量传递方法,用于GEN1对霍利迪结分辨率进行实时调查
DOI:
10.3791/60045-v
•
11:27 min
•
September 18, 2019
•
Mohamed A. Sobhy*
1
,
Amer Bralić*
1
,
Vlad-Stefan Raducanu
,
Muhammad Tehseen
,
Yujing Ouyang
,
Masateru Takahashi
,
Fahad Rashid
,
Manal S. Zaher
,
Samir M. Hamdan
1
Laboratory of DNA Replication and Recombination, Biological and Environmental Sciences and Engineering Division (BESE)
,
King Abdullah University of Science and Technology (KAUST)
Chapters
00:04
Title
01:22
Flow Cell Preparation
03:01
Single-Molecule Förster Resonance Energy Transfer (smFRET) Experiments
06:25
Sm-FRET Data Analysis
07:10
Two-Color Alternating Excitation (ALEX) FRET Experiment and Analysis
08:24
Results: Representative Real-Time HJ Resolution Investigation
10:13
Conclusion
Summary
Automatic Translation
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Automatic Translation
这里介绍的是一个执行单分子Fürster共振能量转移以研究HJ分辨率的协议。双色交替激励用于确定解离常数。然后,在实时裂解测定中应用单色时移smFRET,以获得HJ分辨率前的沉思时间分布。
Tags
Single-molecule
Förster Resonance Energy Transfer (FRET)
Holliday Junction
GEN1
Real-time Investigation
Enzymatic Systems
Single-molecule Methods
Biomolecular Interactions
Optical And Force Measurements
Single-channel Flow Cell
Quartz Slide
Double Adhesive Sheet
Polytetrafluoroethylene Tweezers
Polyethylene Tubing
Epoxy Glue
Avidin
PBS
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