One_Step_Preparation_and_Transformation_of_Competent_Cells

One_Step_Preparation_and_Transformation_of_Competent_Cells...

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One Step Preparation and Transformation of Competent Cells Harman J Singh MBIOTECH 401 10/8/2010
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Abstract Transformation is a very basic technique in molecular biology laboratory. In this technique plasmid is inserted into the bacteria which is then multiplied as the cell divides. In this experiment foreign plasmid was inserted into the E.Coli cells which were made competent by giving them a heat shock to make their cell wall permeable. The resulting cells were grown in the culture media to calculate the transformation efficiency of this procedure. The transformation efficiency came out to be a function of amount of plasmid inserted in the LB media at three different volumes: 10ul; 100ul; 250ul. These came out to be as such 2 × 10^4; 9 × 10^4 & 3.3 × 10^5 transformants/ug for 10ul, 100ul and 250ul aliquots respectively.
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Introduction The principle of transformation was first observed by Fred Griffith in 1928. In doing experiments on mice to determine what strain of Streptococcus pneumonia was responsible for its death, it was observed that the injection of both alive non-lethal cells and heat- killed lethal cells was the cause for the mice death. This lead Fred Griffith to propose that non- lethal cells of the bacteria was transformed by the heat-killed lethal bacteria (Malacinski, 2003). In latter experiments by scientists, it was proved that DNA was the chemical compound that was transformed. Therefore, alternation of DNA or genetic information has ample benefits in modern biotechnology. In addition, the replication ability of bacteria can amplify the production of complex molecules which are then transported into cytoplasm for its various vital functions. The aggregation of bacterial cell is called a colony which can be seen with a naked eye. Bacterial transformation can happen in many ways such as conjugation; transfection and transformation. The purpose of the transformation is to introduce foreign plasmid into a bacteria which then can be used for the amplification to make large quantities of it. The plasmid is a circular DNA found only in bacteria that has a base pair of about 2,000 to 10,000bp. The plasmids have three basic charactertics which is that they contain a selectable marker such as antibiotic gene; an
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One_Step_Preparation_and_Transformation_of_Competent_Cells...

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