Combinatorial DNA taggants and methods of preparation and use thereof
DNA taggants in which the nucleotide sequences are defined according to combinatorial mathematical principles. Methods of defining nucleotide sequences of the combinatorial DNA taggants, and using such taggants for authentication and tracking and tracing an object or process are also disclosed.
1. A method of making a combinatorial DNA taggant comprising:
defining a set of n unique first single stranded DNA strands S 1 through S n and n unique reverse complement strands CS 1 through CS n of the first single stranded DNA strands, wherein n is a natural number having a value greater than 2; and
synthesizing a set of n(n−1)/2 unique bit register single stranded encoding strands comprised of base sequences by:
synthesizing n−1 of the n(n−1)/2 unique bit register single stranded encoding strands, the base sequences of which are synthesized by combining the base sequences of the first of the n unique single stranded DNA strands S 1 with the base sequences of each of reverse complement strand CS 2 through CS n , wherein for each of the n−1 unique bit register single stranded encoding strands, there are no nucleotide bases between the 3′ end of the respective unique single stranded DNA strand and the 5′ end of the unique reverse complement strand; and
synthesizing unique single stranded DNA strands, the base sequences of which are synthesized by combining the base sequences of each of the unique single stranded DNA strands S i through S n−1 , with the base sequences of each of the reverse complement strands CS i+1 through CS n , where i has an initial value of 2, thereby producing the unique bit register single stranded encoding strands, and wherein for each of the unique bit register single stranded encoding strands, there are no nucleotide bases between the 3′ end of the respective unique single stranded DNA strand and the 5′ end of the unique reverse complement strand.
2. The method of claim 1 , further comprising selecting a subset of the n(n−1)/2 unique bit register encoding strands as the combinatorial DNA taggant.
3. The method of claim 2 , wherein the selected subset of the n(n−1)/2 unique bit register encoding strands are mixed into a single liquid solution to make the combinatorial DNA taggant.
4. The method of claim 2 , wherein the n(n−1)/2unique bit register encoding strands are prepared in n(n−1)/2 separate liquid solutions, and the method further comprises mixing the liquid solutions of the selected subset of the n(n−1)/2 unique bit register encoding strands to make the combinatorial DNA taggant.
5. The method of claim 4 , wherein the selected subset of the n(n −1)/2 unique bit register encoding strands are mixed into a single liquid solution to make the combinatorial DNA taggant.