PRIMER EXTENSION MASS SPECTROSCOPY NUCLEIC ACID SEQUENCING METHOD

摘要

A method of sequencing a target nucleic acid using primer extension mass spectroscopy to generate an observed mass spectrum, on which base calling cycles are carried out using calibrated inter-peak mass difference values such that: each called base allows mass calculation for the base-called peak in the observed mass spectrum; this calculated mass is then used as a further calibration point for subsequent rounds of base calling. A reaction mixture of all four base-specific chain extension nucleotides and four chain termination nucleotide analogues. A method of performing mass spectroscopy, which method comprises subjecting molecular ions which have been chemically charged in a predetermined manner to time-of-flight or Fourier transform mass spectroscopy.