Nucleic acid detection method
AssigneeHitachi High-Technologies Corporation
InventorsMaiko Tanabe, Chihiro Uematsu
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Abstract
This invention provides a novel method for amplifying and detecting a target gene rapidly with high sensitivity under isothermal conditions. In such method, a sequence to be amplified can be freely designed regardless of the template sequence, an amplified product can be amplified and detected within a short period of time with high sensitivity, and thus, the gene expression level can be determined more easily than is possible with prior art.
Claims8
A method for detecting a target nucleic acid comprising the following steps, which comprises detecting a sequence that is nonspecific to the target nucleic acid in the transcription product:
- step 1 of subjecting the target nucleic acid as a template to reverse transcription using the first primer comprising the first base sequence complementary to sequence F1 of the target nucleic acid, the second base sequence complementary to a promoter sequence of RNA polymerase, and an arbitrary third base sequence, in that order from the 3′ end;
- step 2 of subjecting the target nucleic acid as a template to digestion with an enzyme;
- step 3 of subjecting the reverse transcription product as a template to elongation using the second primer comprising the fourth base sequence identical to sequence F2 located at a position closer to the 5′ end than the sequence F1 of the target nucleic acid;
- and
- step 4 of subjecting the elongation product to transcription with RNA polymerase.
Dependencies are inferred from the claim text.