FIELD: biotechnology.
SUBSTANCE: for method implementation, a solid substrate is provided on which RNA polymerase is immobilized and sequentially the locations of the first, second and third of the four nucleotides are sequentially detected, and the remaining positions not occupied by the first three are defined as the positions of the fourth nucleotide. To determine each particular position of the first, second and third nucleotide, the RNA polymerase is contacted with a nucleic acid molecule of interest containing a detectable optically rotating label in the first, second and third sequencing conditions, respectively. The first, second and third sequencing conditions involve the presence of four nucleotide triphosphates, wherein the first, second or third nucleotide triphosphate of the four is suitably present in a rate-limiting amount. At that, stopping rotation of the rotating label indicates the presence of the nucleotide triphosphate, which is in the rate-limiting amount.
EFFECT: invention allows to increase the sensitivity and accuracy of quantitative analysis during sequencing.
25 cl, 11 dwg, 5 tbl, 9 ex
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Authors
Dates
2017-11-23—Published
2012-02-23—Filed