大肠杆菌感受态细胞的制备及重组子的转化_实验报告(国外英文资料).docVIP

大肠杆菌感受态细胞的制备及重组子的转化_实验报告(国外英文资料) Molecular biology lab report Experimental name: the preparation of e. coli and the transformation of the recombinant Class: sangon XXXX Name: XXX Student number: XXX Date: XXX The preparation of e. coli and the transformation of the recombinant 1 the introduction Genetic manipulation has become an important conventional technology today in molecular biology. Connection of the recombinant DNA in vitro import appropriate receptor cells can freely copy, proliferation and expression, so as to get a lot of recombinant gene, notably the transformation is given priority to [1]. The preparation of sensory cells is an important part of molecular biology research, and the quality of the preparation is directly influenced by the follow-up experiment. The purpose of this experiment was to understand the concept of, and its significance in molecular biology research, learning methods of competent escherichia coli cells prepared calcium chloride method. 2 materials and methods Experimental principle Plasmid DNA has been conversion process (transformation), in order to import it into cells (competent cell) or e. coli in the body, and then through the host, the system to achieve the purpose of copying DNA of bacteria. For conversion of function one of the most commonly used method is to add a lot of calcium chloride, calcium chloride), lead to structural changes in the cell walls of e. coli, into cells, and is advantageous to the plasmid DNA into bacterial cells. Most strains of e. coli conversion efficiency between 105-108, the per mu g successfully transformed cell number in plasmid DNA, affecting the efficiency factor and cells or the ability to absorb the DNA. And whether the two factors and the bacteria in the logarithmic phase, when the processing is to keep cells at 4 ° C, and calcium chloride processing time of cells. Cells after the completion of the preparation, the use of heat shock treatment, make the cytoplasm membrane fat degeneration