Electrospray Quadrupole Mass Spectrometry Analysis of Model Oligonucleotides and Polymerase Chain Reaction Products: Determination of Base Substitutions, Nucleotide Additions/Deletions, and Chemical M

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• 作者：Mark T. Krahmer ; Yevette A. Johnson ; James J. Walters ; Karen F. Fox ; Alvin Fox ; and Madan Nagpal
• 刊名：Analytical Chemistry
• 出版年：1999
• 出版时间：July 15, 1999
• 年：1999
• 卷：71
• 期：14
• 页码：2893 - 2900
• 全文大小：116K
• 年卷期：v.71,no.14(July 15, 1999)
• ISSN：1520-6882

文摘

ESI FTICR mass spectrometry is the only techniquecurrently used for accurate molecular weight analysis ofPCR products above 100 bp in size. This is important indemonstrating the potential for MS in making majorcontributions in the molecular biology and genomicsareas. In the near future, it is more likely that lessexpensive, more user friendly MS techniques will be usedfor high-throughput analyses (including MALDI TOF andESI quadrupole). There have been numerous reports onthe use of MALDI TOF. The current report is to the firstto evaluate the use of ESI-quadrupole analysis of PCRproducts. Synthetic oligonucleotides (30 and 89 mers)and polymerase chain reaction products of varying molecular weight (62, 88, 89, and 114 bp) were analyzedby ESI using a quadrupole MS. The mass accuracy fornucleic acids in the 30-62 bp range was shown to allowdetermination of nucleotide substitutions and additions/deletions. For higher molecular weight PCR products(88-114 bp), the mass accuracy of ESI-MS distinguishessingle or multiple nucleotide insertions/deletions. Inaddition, ESI quadrupole MS allows determination ofmolecular weight of both strands of higher molecularweight ds PCR products and can distinguish nucleotidemodifications (e.g., with biotin). In conclusion, it isdemonstrated that ESI-MS occupies an intermediate position (as compared to MALDI TOF and ESI FTICR) withregard to mass accuracy and resolution in analysis ofnucleic acids.