Substrate for nucleic acid amplification, and method for manufacturing same
AssigneeKorea Research Institute of Bioscience and Biotechnology
InventorsOh Seok Kwon, Tai Hwan Ha, Kyung Ho Kim, Jin Yeong Kim, Seon Joo Park, Seong Eun Seo
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Abstract
The present invention relates to a substrate for nucleic acid amplification, and a method for manufacturing same, the substrate for rapid and accurate PCR analysis comprising: a transparent substrate; a micro-patterned metal layer formed on the transparent substrate; an N-heterocyclic carbene compound having one end annealed to the surface of the micro-patterned metal layer; and a primer immobilized on the other end of the N-heterocyclic carbene compound.
Claims16
A substrate for nucleic acid amplification, used in light-based polymerase chain reaction (PCR) using thermal energy produced by the irradiation of light energy from light sources, comprising:
- a transparent matrix;
- a micro-patterned metal layer formed on the transparent matrix;
- an N-heterocyclic carbene compound having one end bonded to the surface of the micro-patterned metal layer;
- and
- at least one pair of primers immobilized on the other end of the N-heterocyclic carbene compound,
- wherein the N-heterocyclic carbene compound is represented by the following Chemical Formula 1 or 2:
- in Chemical Formulae 1 and 2,
- R1, R2, R5 and R6 are the same or different, and are each independently hydrogen, an alkyl group of 1 to 20 carbon atoms, a cycloalkyl group of 3 to 20 carbon atoms, an aryl group of 6 to 30 carbon atoms or a heteroaryl group of 2 to 30 carbon atoms,
- R3, R4, R7, R8, R9 and R10 are the same or different, and are each independently, hydrogen, an alkyl group of 1 to 20 carbon atoms, a cycloalkyl group of 3 to 20 carbon atoms, an aryl group of 6 to 30 carbon atoms, a heteroaryl group of 2 to 30 carbon atoms or a structure represented by the following Chemical Formula 3, where adjacent two or more substituents among R7 to R10 are bonded to form a hydrocarbon ring,
- at least one of R3 and R4 is the structure represented by the following Chemical Formula 3, and
- at least one of R7 to R10 is substituted with the structure represented by the following Chemical Formula 3, or in case where adjacent two or more substituents among R7 to R10 are bonded to form a hydrocarbon ring, at least one of carbon atoms forming the hydrocarbon ring is substituted with the structure represented by the following Chemical Formula 3:
- in Chemical Formula 3,
- n is a repeating number of a unit in parenthesis and an integer of 1 to 30, and
- A is a nitrogen (N)-containing alkyl group of 1 to 20 carbon atoms or a nitrogen (N)-containing heteroaryl group of 2 to 30 carbon atoms.
A method for manufacturing a substrate for nucleic acid amplification, wherein the substrate is used in light-based polymerase chain reaction (PCR) using thermal energy produced by the irradiation of light energy from light sources, the method comprising:
- forming a micro-patterned metal layer on a transparent matrix;
- introducing one end of an N-heterocyclic carbene compound onto the surface of the micro-patterned metal layer;
- and
- immobilizing at least one pair of primers on the other end of the N-heterocyclic carbene compound,
- wherein the N-heterocyclic carbene compound is represented by the following Chemical Formula 1 or 2:
- in Chemical Formulae 1 and 2,
- R1, R2, R5 and R6 are the same or different, and are each independently hydrogen, an alkyl group of 1 to 20 carbon atoms, a cycloalkyl group of 3 to 20 carbon atoms, an aryl group of 6 to 30 carbon atoms or a heteroaryl group of 2 to 30 carbon atoms,
- R3, R4, R7, R8, R9 and R10 are the same or different, and are each independently hydrogen, an alkyl group of 1 to 20 carbon atoms, a cycloalkyl group of 3 to 20 carbon atoms, an aryl group of 6 to 30 carbon atoms, a heteroaryl group of 2 to 30 carbon atoms or a structure represented by the following Chemical Formula 3, where adjacent two or more substituents among R7 to R10 are bonded to form a hydrocarbon ring,
- at least one of R3 and R4 is the structure represented by the following Chemical Formula 3, and
- at least one of R7 to R10 is substituted with the structure represented by the following Chemical Formula 3, or in case where adjacent two or more substituents among R7 to R10 are bonded to form a hydrocarbon ring, at least one of carbon atoms forming the hydrocarbon ring is substituted with the structure represented by the following Chemical Formula 3:
- in Chemical Formula 3,
- n is a repeating number of a unit in parenthesis and an integer of 1 to 30, and
- A is a nitrogen (N)-containing alkyl group of 1 to 20 carbon atoms or a nitrogen (N)-containing heteroaryl group of 2 to 30 carbon atoms.
Dependencies are inferred from the claim text.