Nucleic Acids Research, 1974, Vol. 1, No. 11 1503-1520
© 1974
Articles |
DNA sequence analysis: a formula to predict electrophoretic mobilities of oligonucleotides on cellulose acetate*
Section of Biochemistry, Molecular and Cell Biology, Cornell University Ithaca, New York 14850, USA
Received August 19, 1974.
A simple method has recently become available for sequence analysis of large oligonucleotide fragments. Sequences are derived from the characteristic mobility shifts of the sequential partial degradation products of the oligonucleotide on two dimensional homochromatography. We have now developed an empirical formula for predicting the relative mobilities of each of the partial products in the first dimension (electrophoresis on cellulose acetate gel). The formula allows a more precise interpretation of the sequence of the oligonucleotide. It eliminates the ambiguities present in the method previously reported for sequence analysis by simple inspection of the mobility shifts. In order to amplify the mobility shifts so that they may be more easily and accurately measured, methods have been developed for preparing and fractionating the oligonucleotides on 40 x 40 cm DEAE-cellulose plates. Both improvements have proven valuable for direct sequence analysis by mapping.
* This is paper XVIII in a series on NUCLEOTIDE SEQUENCE ANALYSIS OF DNA. Paper XVII is by Ghangas and Wu, J. Biol. Chem. 1974. This work was supported by research grants CA 14989 and GM 13887 from the National Institutes of Health and grant GB 40035 from the National Science Foundation. The authors wish to thank Chen-pei D. Tu and Kenneth Marians for some of the data which appear in table 2.
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