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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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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
Chapters
- 00:05Title
- 01:17Proofreading Reactions
- 03:16Terminate the Reaction and Transfer Reaction Products to a Matrix Chip
- 04:54Setup the Assay Information on the Mass Spectrometry
- 06:18MALDI-TOF Mass Spectrometry (MS) Operation and Data Analysis
- 08:00Results: DNA Proofreading and Repair Analyzed by MALDI-TOF MS
- 09:49Conclusion
A non-labeled, non-radio-isotopic method for DNA polymerase proofreading and a DNA repair assay was developed by using high-resolution MALDI-TOF mass spectrometry and a single nucleotide extension strategy. The assay proved to be very specific, simple, rapid, and easy to perform for proofreading and repair patches shorter than 9-nucleotides.
