CPC C09B 57/02 (2013.01) [C09K 11/06 (2013.01); C12Q 1/6876 (2013.01); C09K 2211/1018 (2013.01)] | 21 Claims |
1. A method of sequencing different template polynucleotides, comprising:
(a) incorporating different types of nucleotides to growing polynucleotide chains in the presence of a DNA polymerase, wherein the growing polynucleotide chains are at least partially complementary to the different template polynucleotides to be sequenced, and wherein each type of nucleotide comprises a 3′ blocking group covalently attached to a 2-deoxyribose sugar of the nucleotide; and
(b) imaging and detecting fluorescent signals from the nucleotides incorporated into the growing polynucleotide chains;
wherein at least one type of nucleotide is labeled with a compound of Formula (I):
wherein:
X is O, S, or NRn, wherein Rn is H, C1-6 alkyl, or C6-10 aryl;
ring A is
R, R1, R2, and R4 are each independently H, halo, —CN, —CO2H, amino, —OH, C-amido, N-amido, —NO2, —SO3H, —SO2NRaRb, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aminoalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;
each R3 is independently —CO2H, —(CH2)p—CO2Rc, —SO3H, or C1-4 alkyl substituted with —CO2H or —SO3H;
p is 1, 2, 3 or 4;
each R5 is independently halo, —CN, —CO2Rf, amino, —OH, C-amido, N-amido, —NO2, —SO3H, —SO2NRaRb, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aminoalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;
each Ra and Rb is independently H or optionally substituted C1-6 alkyl;
each Rc, and Rf is independently H, optionally substituted C1-6 alkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;
m is 0, 1, 2, 3, or 4; and
n is 1, 2 or 3.
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