Method and reagent for gene sequence analysis
AssigneesHitachi, Ltd., National University Corporation Gunma University
InventorsMasayasu Kuwahara, Tomoharu Kajiyama, Hideki Kambara
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Abstract
Provided is a nucleic acid substrate which has nucleic acid substrate characteristics equivalent to those of dATP, has a low substrate specificity for luciferase, exerts no negative effect on enzymatic reactions such as a complementary-strand synthesis, and therefore is particularly suitable for the pyrosequencing method. As a nucleic acid substrate complementary to nucleotide T, a 7-substituted deoxyribonucleotide triphosphate whose 7-position of a purine group is modified by a substituent is used as a substitute for a nucleotide α-thiotriphosphate analog.
Claims13
A method for nucleic acid analysis, comprising the steps of:
- carrying out a complementary-strand synthesis by using a nucleic acid sample as a template and by adding complementary nucleic acid substrates for nucleotides A, G, T, and C;
- generating ATP from pyrophosphate generated in the complementary-strand synthesis by using an enzyme;
- and
- determining the presence or absence of the complementary-strand synthesis by detecting chemiluminescence produced in a luciferase reaction,
- wherein a 7-substituted deoxyribonucleotide triphosphate whose 7-position of a purine group is modified by a substituent is used as the complementary nucleic acid substrate for nucleotide T.
A complementary nucleic acid substrate reagent for nucleotide T, comprising a 7-substituted deoxyribonucleotide triphosphate whose 7-position of a purine group is modified by a substituent.
A compound selected from the following:
A compound represented by the following formula:
Dependencies are inferred from the claim text.