The Experts below are selected from a list of 16965 Experts worldwide ranked by ideXlab platform
Paul D Adams - One of the best experts on this subject based on the ideXlab platform.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
Philip C Gach - One of the best experts on this subject based on the ideXlab platform.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
Jess Sustarich - One of the best experts on this subject based on the ideXlab platform.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
Steve C C Shih - One of the best experts on this subject based on the ideXlab platform.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
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a droplet microfluidic platform for automating genetic engineering
2016Co-Authors: Philip C Gach, Steve C C Shih, Jess Sustarich, Jay D Keasling, Nathan J Hillson, Paul D AdamsAbstract:We present a water-in-oil droplet microfluidic platform for transformation, Culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, Culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the Culture Chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell Culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and Cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.
Ping Wang - One of the best experts on this subject based on the ideXlab platform.
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3d cell based biosensor for cell viability and drug assessment by 3d electric cell matrigel substrate impedance sensing
2019Co-Authors: Xinwei Wei, Li Gong, I Zhang, Ping WangAbstract:Abstract Preclinical efficacy and toxicity assessment of drug candidates plays a significant role in drug discovery and development. Traditional planar cell Culture is a common way to perform the preclinical drug test, but it is difficult to correctly predict the drug efficacy and toxicity due to the simple two-dimensional (2D) extracellular microenvironment. Compared to the planar cell Culture, three-dimensional (3D) cell Culture system can better mimic the complex extracellular microenvironment where cells reside in the 3D tissues/organs in vivo. However, the conventional imaging techniques are difficult to achieve the dynamic and label-free monitoring of cellular behavior in thick sample by 3D cell Culture. Here, 3D electric cell/matrigel-substrate impedance sensing (3D-ECMIS) is developed for real-time and non-invasive monitoring of 3D cell viability and drug susceptibility. In this study, human hepatoma cells (HepG2) are encapsulated in the matrigel scaffold and Cultured in a 3D ECMIS chip which involves a pair of vertical golden electrodes on the opposite sidewalls of the Culture Chamber for the in-situ impedance measurement. Moreover, a portable multichannel system is developed to monitor the 3D cell/matrigel construct. The number of 3D-Cultured cells was inversely proportional to the impedance magnitude of the entire cell/matrigel construct. Furthermore, anti-cancer drug screening will be conducted on the 3D-Cultured cells when the cell proliferation reaches to a plateau phase. To validate the performance of 3D-ECMIS for the cell viability and drug susceptibility, the cell live/dead staining are utilized to confirm the results of drug susceptibility by this 3D-cell-based biosensor system. It is demonstrated that the 3D cell-based biosensor and 3D-ECMIS system will be a promising platform to improve the accuracy of cell-based anti-cancer drug screening in vitro.
