Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

9.5K Views

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11:08 min

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June 19th, 2018

DOI :

10.3791/57862-v

June 19th, 2018

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Kang-Yi Su*¹^,², Steven D. Goodman*³, Hung-Ming Lai¹, Rong-Syuan Yen¹, Wei-Yao Hu¹, Wern-Cherng Cheng², Liang-In Lin¹^,², Ya-Chien Yang¹^,², Woei-Horng Fang¹^,²

¹Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, ²Department of Laboratory Medicine, National Taiwan University Hospital, ³Center for Microbial Pathogenesis, Nationwide Children's Hospital and the Department of Pediatrics, The Ohio State University

Summary

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Keywords DNA Repair

Proofreading

MALDI TOF Mass Spectrometry

Single Nucleotide Extension

DNA Fidelity

DNA Polymerase

Primer template Preparation

Mismatch Substrate

Reaction Buffer

DdNTPs

DNA Polymerase Dilution

Reaction Temperature

Reaction Termination

Chapters in this video

0:05

Title

1:17

Proofreading Reactions

3:16

Terminate the Reaction and Transfer Reaction Products to a Matrix Chip

4:54

Setup the Assay Information on the Mass Spectrometry

6:18

MALDI-TOF Mass Spectrometry (MS) Operation and Data Analysis

8:00

Results: DNA Proofreading and Repair Analyzed by MALDI-TOF MS

9:49

Conclusion

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