Sandbox Reserved 657: Difference between revisions
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== Methods of Analysis == | == Methods of Analysis == | ||
The overall process of analysis of spo0F includes proliferation, isolation, purification, and analysis. One specific method of analysis will be described. The proliferation process can accomplished by transforming competent E. coli cells through heat shock treatment. E. coli is able to relatively easily uptake plasmids and may be induced to over express the protein. The plasmid used in the transformation requires a his-tagged spo0F gene, an antibiotic resistance gene, and a transcription initiation sequence. A common initiation sequence is one that is activated by isopropyl β-D-1-thiogalactopyranoside (IPTG) activation sequence. Once the cells are transformed cellular proliferation is the next step. To isolate the transformed cell, the cells are cultured on a selective medium using the antibiotic that the plasmid provides resistance to. | The overall process of analysis of spo0F includes proliferation, isolation, purification, and analysis. One specific method of analysis will be described. The proliferation process can accomplished by transforming competent E. coli cells through heat shock treatment. E. coli is able to relatively easily uptake plasmids and may be induced to over express the protein. The plasmid used in the transformation requires a his-tagged spo0F gene, an antibiotic resistance gene, and a transcription initiation sequence. A common initiation sequence is one that is activated by isopropyl β-D-1-thiogalactopyranoside (IPTG) activation sequence. Once the cells are transformed cellular proliferation is the next step. To isolate the transformed cell, the cells are cultured on a selective medium using the antibiotic that the plasmid provides resistance to. | ||