GENE CLONING
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Transformation
Transformation is a process by which introduce the vector into bacterial host cells through porous membrane or competent cells.
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Competent cells refer to host cell that undergo specific treatment to increase the transformation efficiency.
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Pores on the membrane should be closed quickly after vector uptake to prevent host cell die
Porous host membrane can be done by:
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Chemical- based treatment
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Treated the cells by calcium chloride and heat shocked at 42 °C for 30 second.
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Advantages: Fast and easy
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Electrical-based treatment (electroporation)
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Electromagnetic field is applied to cells
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Heat shock treatment
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Thaw cells on ice (-80 °C, 20-30 minutes)
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Put the tube into 42 °C water bath for 30-60 seconds.
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Put back the tube into ice for 20 minutes.
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