Difference between revisions of "Team:BostonU HW/Transformation"

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Cell lysis is a commonly used protocol in synthetic biology. It can be performed through a variety of different methods, however we had focused on chemical cellular lysis. Cell Lysis is used to extract and isolate DNA from a specific type of cell. This is an extremely important step in building genetic circuits in order to utilize specific coding regions in a cell's DNA. Chemical cell lysis involves introducing cells to a series of buffers in order to degrade the cell’s outer membrane and collect the DNA that is released.
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Bacterial transformation is a commonly used protocol in synthetic biology. It can be used for a variety of functions, such as testing whether or not a genetic circuit is functional. Transformation allows bacterial cells, such as Escherichia coli, to take in and express external DNA fragments. Transformation consists of heat shock to damage cells and promote the taking up of external plasmids, recovery to prevent cells from dying, and a final culturing. From there, cells are analyzed.
 
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This microfluidic chip is designed to perform chemical cell lysis. Suspended cells and an lysing buffer would be mixed inside the cell. This mixture of buffer and cells would be then mixed with a neutralization buffer to prevent DNA degradation. The mixture would then move to the diamond chamber where the DNA binds magnetic particles. Lastly, an elution buffer would be input and to clean and release the DNA from the magnetic particles.
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This microfluidic chip is designed to perform transformation. Suspended cells and plasmid are metered on the chip and are then mixed together. The solution then undergoes heat shock in a time-dependent mixing element for exactly 30 seconds. The solution can then be pipetted out from the chip into a recovery tube on ice.
 
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Revision as of 05:15, 29 October 2017

BostonU_HW

Cell Lysis

Summary

Bacterial transformation is a commonly used protocol in synthetic biology. It can be used for a variety of functions, such as testing whether or not a genetic circuit is functional. Transformation allows bacterial cells, such as Escherichia coli, to take in and express external DNA fragments. Transformation consists of heat shock to damage cells and promote the taking up of external plasmids, recovery to prevent cells from dying, and a final culturing. From there, cells are analyzed.
This microfluidic chip is designed to perform transformation. Suspended cells and plasmid are metered on the chip and are then mixed together. The solution then undergoes heat shock in a time-dependent mixing element for exactly 30 seconds. The solution can then be pipetted out from the chip into a recovery tube on ice.
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Testing

This following video shows a test of the chip using colored water. This is to show help explain the functionality of the chip. No biological material was inserted into this chip.