Molecular weight determination of plasmid DNA using electrospray ionization mass spectrometry

被引:67
作者
Cheng, XH
Camp, DG
Wu, QY
Bakhtiar, R
Springer, DL
Morris, BJ
Bruce, JE
Anderson, GA
Edmonds, CG
Smith, RD
机构
[1] PACIFIC NW LAB, ENVIRONM MOL SCI LAB, RICHLAND, WA 99352 USA
[2] EASTERN OREGON STATE COLL, DEPT CHEM, LA GRANDE, OR 97850 USA
[3] PACIFIC NW LAB, DEPT BIOL & CHEM, RICHLAND, WA 99352 USA
关键词
D O I
10.1093/nar/24.11.2183
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
lonization and molecular weight (MW) determination of megadalton size plasmid DNA has been achieved using electrospray ionization (ESI) with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry, DNA molecules were shown to remain intact through electrospray ionization by collection on a specially prepared surface, followed by agarose gel electrophoresis. Individual highly charged ions of plasmid DNA produced by ESI were trapped in an FTICR cell for up to several hours and reacted with acetic acid to induce charge state shifts, Measurements of mass-to-charge ratios for these multiple peaks arising from charge state shifting give MW measurements of individual ions with an average accuracy of 0.2%, The MW distribution was obtained by measurements for a number of individual ions from the same sample [plasmid DNA: pGEM-5S MW(cal) = 1.946 MDa], yielding a MW(obs) of 1.95 +/- 0.07 MDa for ions clustered in the vicinity of the expected MW.
引用
收藏
页码:2183 / 2189
页数:7
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