This paper represents an early work towards the simplification of DNA sequencing, in the time when the Sanger dideoxy method was not yet quite established. The authors found modified chemical conditions, whereby chemical degradation of each of the four bases was effected in such a way that differential intensities were found that allowed for correct base calling. All oligos were labeled using T4polynucleotide kinase and P32. The chemical degradation conditions were: 0.4 aqueous piperidinium formate, pH 2, at 37C for 6 h, followed by treatment with 1 aqueous piperidine at 90C for 6 h. The degradation products were analyzed in a 27% polyacrylamide gel; the differential intensities of each of the bases was found to be G > A > C > T. Also the spacing obtained from a given band in relation to the next higher band in the ladder was unique for each base , this characteristic order of spacing was found to be G > T A > C. This method is an improvement of the previous report by one of the authors, since this procedure renders a salt-free hydrolyzate, in such a way as to minimize possible electrophoretic artifacts of the smaller bands running at the front of the PAGE gel.
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