Abstract
On microfluidic chips, aqueous droplets are generated by simultaneously injecting aqueous phase and oil phase into the microfluidic channels at controlled flow rates. Aqueous droplets surrounded by oil offers a simple solution to confine the dispersion of reagents, so that biological and chemical reactions can be controlled precisely. In this protocol, one of the common yeast cell transformation approaches, electroporation, is successfully performed on chip by manipulating aqueous droplets with an electrochemical detection method. Yeast cells in aqueous droplet are electroporated by applying AC voltage at 18 Vpp (peak-to-peak) and 1 kHz. The whole experimental flow consists of masks design, microelectrode fabrication, microfluidic channel fabrication, and yeast cell electroporation.
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- 1.
Latex hampers PDMS curing, so you’d better not wear latex gloves when handling non-cured PDMS.
- 2.
When using a negative photoresist such as SU-8, the dark and light region of photomask should be designed in an opposite way.
- 3.
For a 4-in. wafer 50 g PDMS with 5 g curing agent should be enough. If your wafer size is different, adjust the amount of PDMS accordingly.
- 4.
To prevent liquid leakage from the inlet and outlet, the diameter of the puncher should be a little smaller than that of the tubing.
- 5.
Since plasma bonding is irreversible, don’t attach the PDMS to the slide until they are well aligned.
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For transformation and transfection process, fluorescein can be changed to customized target plasmids or DNA sequences.
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© 2015 Springer International Publishing Switzerland
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Cai, Q., Luo, C. (2015). Yeast Cell Electroporation in Droplet-Based Microfluidic Chip. In: van den Berg, M., Maruthachalam, K. (eds) Genetic Transformation Systems in Fungi, Volume 2. Fungal Biology. Springer, Cham. https://doi.org/10.1007/978-3-319-10503-1_18
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DOI: https://doi.org/10.1007/978-3-319-10503-1_18
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Publisher Name: Springer, Cham
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Online ISBN: 978-3-319-10503-1
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