The conditions under which oligonucleotides hybridize only with entirely
homologous sequences are recognized. The sequence of a given
DNA fragment is read by the hybridization and
assembly of positively hybridizing probes through overlapping portions. By simultaneous hybridization of
DNA molecules applied as dots and bound onto a filter, representing single-stranded phage vector with the cloned insert, with about 50,000 to 100,000 groups of probes, the main type of which is (A,T,C,G)(A,T,C,G)N8(A,T,C,G), information for computer determination of a sequence of
DNA having the complexity of a
mammalian genome are obtained in one step. To obtain a maximally completed sequence, three libraries are cloned into the phage vector, M13,
bacteriophage are used: with the 0.5 kb and 7 kbp insert consisting of two sequences, with the average distance in
genomic DNA of 100 kbp. For a million bp of
genomic DNA, 25,000 subclones of the 0.5 kbp are required as well as 700 subclones 7 kb long and 170
jumping subclones. Subclones of 0.5 kb are applied on a filter in groups of 20 each, so that the total number of samples is 2,120 per million bp. The process can be easily and entirely robotized for factory reading of complex genomic fragments or DNA molecules.