Matrix-assisted Laser Desorption/Ionization for Sequencing Single-stranded and Double-stranded DNA

✍ Scribed by N. I. Taranenko; C. N. Chung; Y. F. Zhu; S. L. Allman; V. V. Golovlev; N. R. Isola; S. A. Martin; L. A. Haff; C. H. Chen

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 230 KB
• Volume: 11
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(19970228)11:4<386::aid-rcm867>3.0.co;2-n

✦ Synopsis

The DNA sequence of a single-stranded and double-stranded template was determined. The templates were sequenced using the chain termination method and cycle sequencing method and detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The sequencing products were analyzed successfully without the laborious and expensive methods for removal of the template. Direct sequencing of the double-stranded template was achieved with minimal post-reaction purifications, which could be extremely important for mutation analysis and clinical diagnosis. A systematic study of the mechanisms and kinetics of sequencing reactions was also performed. The details of this analysis and directions for future improvements of the quality of sequencing are presented.