Dna sequencing and dna terminators

C - Chemistry – Metallurgy – 12 – P

Patent

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C12P 19/34 (2006.01) C07H 19/044 (2006.01) C07H 19/20 (2006.01) C09B 23/12 (2006.01) C12Q 1/68 (2006.01) G01N 27/447 (2006.01)

Patent

CA 2181219

A universal terminator includes a heterocycle other than naturally occuring DNA heterocycles such as adenine, cytosine, guanine and thymine. In terminating strand synthesis, the hybridized primer-template is split into four aliquots. In the "A" vial is added DNA polymerase and normal amounts of "C", "G", and "T" deoxynucleotides, along with a reduced amount of "A" deoxynucleotide and an amount of the universal terminator such that statistically the universal terminator has less than a one percent chance of being incorporated at sites where a "C", "G", or "T" deoxynucleotide should be incorporated and about a one percent chance of being incorporated at sites where an "A" deoxynucleotide should be incorporated. Similar strategies are followed for the "C", "G", and "T" vials, except that the amount of "C", "G" and "T" deoxynucleotides are reduced for their respective vial.

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