Method of making double-stranded DNA sequences

A method of characterizing double-stranded DNA sequences, comprising the steps of:

• (a) providing a double-stranded DNA sequence to be characterized;
• (b) preparing a first series of oligodeoxyribonucleotide fragments which, when joined in proper sequence, yield the expected coding strand of the double-stranded DNA sequence;
• (c) preparing a second series of oligodeoxyribonucleotide fragments which, when joined in proper sequence, yield a DNA strand complementary to the coding strand;
• (d) arbitrarily selecting a first primer for the expected coding strand and a second primer for the complementary strand;
• (e) treating the double-stranded DNA sequence with one oligonucleotide primer for each strand under hybridizing condition;
• (f) polymerizing an extension product of each primer that is complementary to each strand of the double-stranded DNA sequence which is a template for forming the primer extension product;
• (g) denaturing the product of step (f) to separate the primer extension products from their respective templates to form four separate single stranded DNA sequences;
• (h) treating the denatured product of (g) with oligonucleotide primers, such that a primer extension product is synthesizing using each of the single strands produced in step (g) as a template resulting in amplification of the double-stranded DNA sequence;
• (i) repeating steps (e), (f), (g) and (h) until the desired quantity of the double-stranded DNA sequence is formed;
• (j) determining whether the primer extension products are of the expected size and composition by comparison to a standard have the expected size and composition;
• and
• (k) repeating steps (a)-(j) for each remaining oligodeoxyribonucleotide fragment prepared in steps (b) and (c).