A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip ...A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip is simulated and analyzed.The optimal size of the chip is 30×22 mm2, the roundabout micro-channel is the 90 μm width, 40 μm depth. Two micro-heater with the nickel-chrome alloy material film are formed on the side of silicon belonging to denaturation and renaturation zones needed for PCR reaction, and two adiabatic structures with groove on side of silicon by anisotropy etching. By the mode of heating local zones at single side, three wider constant temperature zones could be formed, which are 60 ℃,72 ℃,95 ℃ and suitable for PCR,and the temperature-difference could be restricted in less than 5 ℃.展开更多
Hypertrophic cardiomyopathy (HCM) is one of the diseases damaging people health most badly and some mutations of exons in cardiac troponin I (cTnI) gene are closely associated with family hypertrophic cardiomyopathy (...Hypertrophic cardiomyopathy (HCM) is one of the diseases damaging people health most badly and some mutations of exons in cardiac troponin I (cTnI) gene are closely associated with family hypertrophic cardiomyopathy (FHCM).A microarray was fabricated to screen mutations in exons 3,5,7,and 8 in cTnI gene.Primers were designed for the PCR (polymerase chain reaction) to amplify the target DNA fragments from fresh blood samples.In order to simplify the PCR process,multiplex PCR technology was investigated in detail.The concentration of Mg^(2+) played an important role in multiplex PCR process,a properly low concentration of Mg^(2+) submitted a better speciality of PCR products.The speciality was also favored when the annealing temperature was reasonably enhanced and 64℃is the optimal annealing temperature for the multiplex PCR systems.When applying the fabricated gene-chip to detect the target fragments from PCR mixture,the signal intensity sequence is in accordance with that from theoretic estimate.展开更多
基金the National Natural Science Foundation of China(Grant No.60576047)
文摘A novel continuous-flow PCR chip adopting self-heating, passive-cooling mode to realize the DNA fragments amplification was presented. Using the ANSYS finite element analysis, the temperature distribution of the chip is simulated and analyzed.The optimal size of the chip is 30×22 mm2, the roundabout micro-channel is the 90 μm width, 40 μm depth. Two micro-heater with the nickel-chrome alloy material film are formed on the side of silicon belonging to denaturation and renaturation zones needed for PCR reaction, and two adiabatic structures with groove on side of silicon by anisotropy etching. By the mode of heating local zones at single side, three wider constant temperature zones could be formed, which are 60 ℃,72 ℃,95 ℃ and suitable for PCR,and the temperature-difference could be restricted in less than 5 ℃.
文摘Hypertrophic cardiomyopathy (HCM) is one of the diseases damaging people health most badly and some mutations of exons in cardiac troponin I (cTnI) gene are closely associated with family hypertrophic cardiomyopathy (FHCM).A microarray was fabricated to screen mutations in exons 3,5,7,and 8 in cTnI gene.Primers were designed for the PCR (polymerase chain reaction) to amplify the target DNA fragments from fresh blood samples.In order to simplify the PCR process,multiplex PCR technology was investigated in detail.The concentration of Mg^(2+) played an important role in multiplex PCR process,a properly low concentration of Mg^(2+) submitted a better speciality of PCR products.The speciality was also favored when the annealing temperature was reasonably enhanced and 64℃is the optimal annealing temperature for the multiplex PCR systems.When applying the fabricated gene-chip to detect the target fragments from PCR mixture,the signal intensity sequence is in accordance with that from theoretic estimate.
