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Abstract

A nucleic acid construct useful in preparing reagents for determining target nucleotide sequences in the nucleic acid of a biological sample, the construct having in its single-stranded form: (a) a target binding region substantially complementary to the target nucleotide sequence, and (b) a signal strand pairing segment bound in the construct by complementary base pairing to a portion of the target binding region; a second portion of the target binding region being single-stranded; and the target binding region and signal strand pairing segment being covalently linked by a phosphate/sugar backbone. A replicable nucleic acid having an origin of replication and two half-restriction sites capable of forming a restriction site can be treated with a restriction enzyme to form a length of nucleic acid containing the target binding region and the signal strand pairing segment. Subsequent labeling of the construct and various optional cleavage and derivation steps can convert the construct to a reagent complex.

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First Claim