Compare calcium chloride transformation, electroporation and transfection. State the benefits and drawbacks of each method and describe...

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Biology

Compare calcium chloride transformation, electroporation andtransfection. State the benefits and drawbacks of each method anddescribe the situations in which each method may be used.
What are the challenges of using biolistics for geneticallymodified organisms? Give one case study on the use ofbiolistics.
Explain the use of Agrobacterium tumerfaciens in geneticallymodifying crops?

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Calcium chloride transformation Competent cells are bacterial cells that can accept extrachromosomal DNA or plasmids naked DNA from the environment The generation of competent cells may occur through two methods natural competence and artificial competence Natural competence is the genetic ability of a bacterium to receive environmental DNA under natural or in vitro conditions Bacteria can also be made competent artificially by chemical treatment and heat shock to make them transiently permeable to DNA This process of making bacterial cells competent by making small holes in the bacterial cell is done using calcium chloride The calcium chloride transformation technique is the most efficient technique among competent cell preparation protocols It increases the bacterial cells ability to incorporate plasmid DNA facilitating genetic transformation The addition of calcium chloride to the cell suspension allows the binding of plasmid DNA to LPS Thus both the negatively charged DNA backbone and LPS come together and when heat shock is provided plasmid DNA passes into the bacterial cell Advantage and disadvantages The cost efficiency and simplicity of the method allows its use in many different cultures this method can accommodate a high concentration of DNA One of the drawbacks is that the Calcium chloride method has only 10 efficiency Also sometimes the calcium chloride can disrupt the cell membrane and cause cell death Thus it is not suitable for protocols that See Answer

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