MALDI-TOF mass spectrometric typing of single nucleotide polymorphisms with mass-tagged ddNTPs.
A matrix-assisted laser desorption/ionization time-of-flight mass spectrometry based method has recently been reported for the typing of single nucleotide polymorphisms using single nucleotide primer extension. This method is limited in some cases by the resolution of the mass determination, as the mass difference between nucleotides can be as little as 9 Da (the difference between A and T). A variation of this method is described here in which a mass-tagged dideoxynucleotide is employed in the primer extension reactions in place of the unmodified dideoxynucleotide. The increased mass difference due to the presence of the mass-tags substantially improves the accuracy and versatility of the procedure.
Fei
Z
Z
Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, WI 53706-1396, USA. fei@whitewater.chem.wisc.edu
Ono
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T
Smith
L M
LM
D015224:Dideoxynucleosides / Q000032:analysis; D009711:Nucleotides; D009838:Oligodeoxyribonucleotides / Q000032:analysis; D011110:Polymorphism, Genetic; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D015224:Dideoxynucleosides; D009711:Nucleotides; D009838:Oligodeoxyribonucleotides