A versatile and simplified non-random strategy for nucleotide sequencing.

We describe a versatile and simplified non-random strategy for nucleotide sequencing. This procedure avoids exposure of the vector DNA to endo- or exonucleases during construction of the subfragment library. The advantages resulting from this strategy are: (1) minimal manipulation in construction of the subfragment library, (2) no need for compatible restriction sites, (3) no insert size limitation, (4) can be used with both chemical and enzymatic sequencing methods. Hence, the procedure provides simplicity, universality and versatility for non-random nucleotide sequencing. We demonstrate the usefulness of this procedure by obtaining the nucleotide sequence of a goat 3.7-kb EcoRI fragment, which is 5' of the epsilon globin gene.