The Experts below are selected from a list of 213 Experts worldwide ranked by ideXlab platform

Michael C Murphy - One of the best experts on this subject based on the ideXlab platform.

  • titer plate formatted continuous flow Thermal Reactors design and performance of a nanoliter reactor
    Sensors and Actuators B-chemical, 2010
    Co-Authors: Pin-chuan Chen, Daniel Sang Won Park, Dimitris E Nikitopoulos, Taehyun Park, Steve Allan Soper, Michael C Murphy
    Abstract:

    Abstract Arrays of continuous flow Thermal Reactors were designed, configured, and fabricated in a 96-device (12 × 8) titer-plate format with overall dimensions of 120 mm × 96 mm, with each reactor confined to a 8 mm × 8 mm footprint. To demonstrate the potential, individual 20-cycle (740 nl) and 25-cycle (990 nl) Reactors were used to perform the continuous flow polymerase chain reaction (CFPCR) for amplification of DNA fragments of different lengths. Since Thermal isolation of the required temperature zones was essential for optimal biochemical reactions, three finite element models, executed with ANSYS (v. 11.0, Canonsburg, PA), were used to characterize the Thermal performance and guide system design: (1) a single device to determine the dimensions of the Thermal management structures; (2) a single CFPCR device within an 8 mm × 8 mm area to evaluate the integrity of the thermostatic zones; and (3) a single, straight microchannel representing a single loop of the spiral CFPCR device, accounting for all of the heat transfer modes, to determine whether the PCR cocktail was exposed to the proper temperature cycling. In prior work on larger footprint devices, simple grooves between temperature zones provided sufficient Thermal resistance between zones. For the small footprint reactor array, 0.4 mm wide and 1.2 mm high fins were necessary within the groove to cool the PCR cocktail efficiently, with a temperature gradient of 15.8 °C/mm, as it flowed from the denaturation zone to the renaturation zone. With temperature tolerance bands of ±2 °C defined about the nominal temperatures, more than 72.5% of the microchannel length was located within the desired temperature bands. The residence time of the PCR cocktail in each temperature zone decreased and the transition times between zones increased at higher PCR cocktail flow velocities, leading to less time for the amplification reactions. Experiments demonstrated the performance of the CFPCR devices as a function of flow velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 min for 20 cycles (24.48 s/cycle) to 2.72 min for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench-top Thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow Thermal Reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer-plate and is the smallest continuous flow PCR to date.

Pin-chuan Chen - One of the best experts on this subject based on the ideXlab platform.

  • titer plate formatted continuous flow Thermal Reactors design and performance of a nanoliter reactor
    Sensors and Actuators B-chemical, 2010
    Co-Authors: Pin-chuan Chen, Daniel Sang Won Park, Dimitris E Nikitopoulos, Taehyun Park, Steve Allan Soper, Michael C Murphy
    Abstract:

    Abstract Arrays of continuous flow Thermal Reactors were designed, configured, and fabricated in a 96-device (12 × 8) titer-plate format with overall dimensions of 120 mm × 96 mm, with each reactor confined to a 8 mm × 8 mm footprint. To demonstrate the potential, individual 20-cycle (740 nl) and 25-cycle (990 nl) Reactors were used to perform the continuous flow polymerase chain reaction (CFPCR) for amplification of DNA fragments of different lengths. Since Thermal isolation of the required temperature zones was essential for optimal biochemical reactions, three finite element models, executed with ANSYS (v. 11.0, Canonsburg, PA), were used to characterize the Thermal performance and guide system design: (1) a single device to determine the dimensions of the Thermal management structures; (2) a single CFPCR device within an 8 mm × 8 mm area to evaluate the integrity of the thermostatic zones; and (3) a single, straight microchannel representing a single loop of the spiral CFPCR device, accounting for all of the heat transfer modes, to determine whether the PCR cocktail was exposed to the proper temperature cycling. In prior work on larger footprint devices, simple grooves between temperature zones provided sufficient Thermal resistance between zones. For the small footprint reactor array, 0.4 mm wide and 1.2 mm high fins were necessary within the groove to cool the PCR cocktail efficiently, with a temperature gradient of 15.8 °C/mm, as it flowed from the denaturation zone to the renaturation zone. With temperature tolerance bands of ±2 °C defined about the nominal temperatures, more than 72.5% of the microchannel length was located within the desired temperature bands. The residence time of the PCR cocktail in each temperature zone decreased and the transition times between zones increased at higher PCR cocktail flow velocities, leading to less time for the amplification reactions. Experiments demonstrated the performance of the CFPCR devices as a function of flow velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 min for 20 cycles (24.48 s/cycle) to 2.72 min for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench-top Thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow Thermal Reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer-plate and is the smallest continuous flow PCR to date.

  • Titer plate formatted continuous flow Thermal Reactors for high throughput applications: Fabrication and testing
    Journal of Micromechanics and Microengineering, 2010
    Co-Authors: Daniel Sang Won Park, Proyag Datta, Namwon Kim, Tae Yoon Lee, Byoung Hee You, Yonas Desta, Taehyun Park, Pin-chuan Chen, Steve Allan Soper, Dimitris E Nikitopoulos
    Abstract:

    A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm 2 , matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow Thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during Thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the Thermal Reactors and the opposite side was patterned with structures to enhance Thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s −1 (15.3 s cycle −1 ) and a relatively low amplification efficiency compared to a bench-top Thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s −1 (46.2 s cycle −1 ) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.

Dimitris E Nikitopoulos - One of the best experts on this subject based on the ideXlab platform.

  • titer plate formatted continuous flow Thermal Reactors design and performance of a nanoliter reactor
    Sensors and Actuators B-chemical, 2010
    Co-Authors: Pin-chuan Chen, Daniel Sang Won Park, Dimitris E Nikitopoulos, Taehyun Park, Steve Allan Soper, Michael C Murphy
    Abstract:

    Abstract Arrays of continuous flow Thermal Reactors were designed, configured, and fabricated in a 96-device (12 × 8) titer-plate format with overall dimensions of 120 mm × 96 mm, with each reactor confined to a 8 mm × 8 mm footprint. To demonstrate the potential, individual 20-cycle (740 nl) and 25-cycle (990 nl) Reactors were used to perform the continuous flow polymerase chain reaction (CFPCR) for amplification of DNA fragments of different lengths. Since Thermal isolation of the required temperature zones was essential for optimal biochemical reactions, three finite element models, executed with ANSYS (v. 11.0, Canonsburg, PA), were used to characterize the Thermal performance and guide system design: (1) a single device to determine the dimensions of the Thermal management structures; (2) a single CFPCR device within an 8 mm × 8 mm area to evaluate the integrity of the thermostatic zones; and (3) a single, straight microchannel representing a single loop of the spiral CFPCR device, accounting for all of the heat transfer modes, to determine whether the PCR cocktail was exposed to the proper temperature cycling. In prior work on larger footprint devices, simple grooves between temperature zones provided sufficient Thermal resistance between zones. For the small footprint reactor array, 0.4 mm wide and 1.2 mm high fins were necessary within the groove to cool the PCR cocktail efficiently, with a temperature gradient of 15.8 °C/mm, as it flowed from the denaturation zone to the renaturation zone. With temperature tolerance bands of ±2 °C defined about the nominal temperatures, more than 72.5% of the microchannel length was located within the desired temperature bands. The residence time of the PCR cocktail in each temperature zone decreased and the transition times between zones increased at higher PCR cocktail flow velocities, leading to less time for the amplification reactions. Experiments demonstrated the performance of the CFPCR devices as a function of flow velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 min for 20 cycles (24.48 s/cycle) to 2.72 min for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench-top Thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow Thermal Reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer-plate and is the smallest continuous flow PCR to date.

  • Titer plate formatted continuous flow Thermal Reactors for high throughput applications: Fabrication and testing
    Journal of Micromechanics and Microengineering, 2010
    Co-Authors: Daniel Sang Won Park, Proyag Datta, Namwon Kim, Tae Yoon Lee, Byoung Hee You, Yonas Desta, Taehyun Park, Pin-chuan Chen, Steve Allan Soper, Dimitris E Nikitopoulos
    Abstract:

    A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm 2 , matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow Thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during Thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the Thermal Reactors and the opposite side was patterned with structures to enhance Thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s −1 (15.3 s cycle −1 ) and a relatively low amplification efficiency compared to a bench-top Thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s −1 (46.2 s cycle −1 ) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.

Daniel Sang Won Park - One of the best experts on this subject based on the ideXlab platform.

  • titer plate formatted continuous flow Thermal Reactors design and performance of a nanoliter reactor
    Sensors and Actuators B-chemical, 2010
    Co-Authors: Pin-chuan Chen, Daniel Sang Won Park, Dimitris E Nikitopoulos, Taehyun Park, Steve Allan Soper, Michael C Murphy
    Abstract:

    Abstract Arrays of continuous flow Thermal Reactors were designed, configured, and fabricated in a 96-device (12 × 8) titer-plate format with overall dimensions of 120 mm × 96 mm, with each reactor confined to a 8 mm × 8 mm footprint. To demonstrate the potential, individual 20-cycle (740 nl) and 25-cycle (990 nl) Reactors were used to perform the continuous flow polymerase chain reaction (CFPCR) for amplification of DNA fragments of different lengths. Since Thermal isolation of the required temperature zones was essential for optimal biochemical reactions, three finite element models, executed with ANSYS (v. 11.0, Canonsburg, PA), were used to characterize the Thermal performance and guide system design: (1) a single device to determine the dimensions of the Thermal management structures; (2) a single CFPCR device within an 8 mm × 8 mm area to evaluate the integrity of the thermostatic zones; and (3) a single, straight microchannel representing a single loop of the spiral CFPCR device, accounting for all of the heat transfer modes, to determine whether the PCR cocktail was exposed to the proper temperature cycling. In prior work on larger footprint devices, simple grooves between temperature zones provided sufficient Thermal resistance between zones. For the small footprint reactor array, 0.4 mm wide and 1.2 mm high fins were necessary within the groove to cool the PCR cocktail efficiently, with a temperature gradient of 15.8 °C/mm, as it flowed from the denaturation zone to the renaturation zone. With temperature tolerance bands of ±2 °C defined about the nominal temperatures, more than 72.5% of the microchannel length was located within the desired temperature bands. The residence time of the PCR cocktail in each temperature zone decreased and the transition times between zones increased at higher PCR cocktail flow velocities, leading to less time for the amplification reactions. Experiments demonstrated the performance of the CFPCR devices as a function of flow velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 min for 20 cycles (24.48 s/cycle) to 2.72 min for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench-top Thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow Thermal Reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer-plate and is the smallest continuous flow PCR to date.

  • Titer plate formatted continuous flow Thermal Reactors for high throughput applications: Fabrication and testing
    Journal of Micromechanics and Microengineering, 2010
    Co-Authors: Daniel Sang Won Park, Proyag Datta, Namwon Kim, Tae Yoon Lee, Byoung Hee You, Yonas Desta, Taehyun Park, Pin-chuan Chen, Steve Allan Soper, Dimitris E Nikitopoulos
    Abstract:

    A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm 2 , matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow Thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during Thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the Thermal Reactors and the opposite side was patterned with structures to enhance Thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s −1 (15.3 s cycle −1 ) and a relatively low amplification efficiency compared to a bench-top Thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s −1 (46.2 s cycle −1 ) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.

Steve Allan Soper - One of the best experts on this subject based on the ideXlab platform.

  • titer plate formatted continuous flow Thermal Reactors design and performance of a nanoliter reactor
    Sensors and Actuators B-chemical, 2010
    Co-Authors: Pin-chuan Chen, Daniel Sang Won Park, Dimitris E Nikitopoulos, Taehyun Park, Steve Allan Soper, Michael C Murphy
    Abstract:

    Abstract Arrays of continuous flow Thermal Reactors were designed, configured, and fabricated in a 96-device (12 × 8) titer-plate format with overall dimensions of 120 mm × 96 mm, with each reactor confined to a 8 mm × 8 mm footprint. To demonstrate the potential, individual 20-cycle (740 nl) and 25-cycle (990 nl) Reactors were used to perform the continuous flow polymerase chain reaction (CFPCR) for amplification of DNA fragments of different lengths. Since Thermal isolation of the required temperature zones was essential for optimal biochemical reactions, three finite element models, executed with ANSYS (v. 11.0, Canonsburg, PA), were used to characterize the Thermal performance and guide system design: (1) a single device to determine the dimensions of the Thermal management structures; (2) a single CFPCR device within an 8 mm × 8 mm area to evaluate the integrity of the thermostatic zones; and (3) a single, straight microchannel representing a single loop of the spiral CFPCR device, accounting for all of the heat transfer modes, to determine whether the PCR cocktail was exposed to the proper temperature cycling. In prior work on larger footprint devices, simple grooves between temperature zones provided sufficient Thermal resistance between zones. For the small footprint reactor array, 0.4 mm wide and 1.2 mm high fins were necessary within the groove to cool the PCR cocktail efficiently, with a temperature gradient of 15.8 °C/mm, as it flowed from the denaturation zone to the renaturation zone. With temperature tolerance bands of ±2 °C defined about the nominal temperatures, more than 72.5% of the microchannel length was located within the desired temperature bands. The residence time of the PCR cocktail in each temperature zone decreased and the transition times between zones increased at higher PCR cocktail flow velocities, leading to less time for the amplification reactions. Experiments demonstrated the performance of the CFPCR devices as a function of flow velocity, fragment length, and copy number. A 99 bp DNA fragment was successfully amplified at flow velocities from 1 mm/s to 3 mm/s, requiring from 8.16 min for 20 cycles (24.48 s/cycle) to 2.72 min for 20 cycles (8.16 s/cycle), respectively. Yield compared to the same amplification sequence performed using a bench-top Thermal cycler decreased nonlinearly from 73% (at 1 mm/s) to 13% (at 3 mm/s) with shorter residence time at the optimal temperatures for the reactions due to increased flow rate primarily responsible. Six different DNA fragments with lengths between 99 bp and 997 bp were successfully amplified at 1 mm/s. Repeatable, successful amplification of a 99 bp fragment was achieved with a minimum of 8000 copies of the DNA template. This is the first demonstration and characterization of continuous flow Thermal Reactors within the 8 mm × 8 mm footprint of a 96-well micro-titer-plate and is the smallest continuous flow PCR to date.

  • Titer plate formatted continuous flow Thermal Reactors for high throughput applications: Fabrication and testing
    Journal of Micromechanics and Microengineering, 2010
    Co-Authors: Daniel Sang Won Park, Proyag Datta, Namwon Kim, Tae Yoon Lee, Byoung Hee You, Yonas Desta, Taehyun Park, Pin-chuan Chen, Steve Allan Soper, Dimitris E Nikitopoulos
    Abstract:

    A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm 2 , matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow Thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during Thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the Thermal Reactors and the opposite side was patterned with structures to enhance Thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s −1 (15.3 s cycle −1 ) and a relatively low amplification efficiency compared to a bench-top Thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s −1 (46.2 s cycle −1 ) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.