Genetic analysis protocols

It is indispensable for any formal genetic analysis to be able to isolate mutants defective in genes coding for important cellular functions. In addition to the spontaneous mutations mentioned above, as caused by ISH elements, several protocols of induced mutagenesis were introduced. The most efficient method so far is the use of the alkylating agent ethyl methane sulfonate to cause point mutations... 

The author would like to thank Dr. Victor Veculescu at Johns Hopkins for development of the sequencing protocols for using SpectruMedix SCE9610 Genetic Analysis System and Daniel Broderick for his thoughtful edits. 

Todd, R.B., Davis, M.A., Hynes, M.J., 2007a. Genetic manipulation olAspergilus nidulans meiotic progeny for genetic analysis and strain construction. Nature Protocols 2,811-821. 

Raw data generated by the Phenotype Pfinder protocol is maintained in a dedicated database. The database automatically graphs the data, generates statistical analyses of assays, and flags those assays which show a statistically significant difference between WT and KO mice. The data can be searched and viewed by either specific KO line or assay. The database also offers a tool for analysis of historically accumulated control data sorted by genetic background. 

Any document used to collect research data on clinical study subjects may be genetically classed as a data collection form. These completed forms provide evidence of the research conducted. The most common type of data collection form is the CRF. Other types of data collection forms include diary cards, dispensing records, quality of life forms, etc. The CRF must allow for proper analysis of the data and proper reporting of the data in the final clinical study report and it must reflect the protocol exactly no more and no less data must be collected. Thus, a CRF must be created for each clinical study and must be prepared in parallel with the protocol. CRFs are usually also prepared by spon-sors/CROs in pharmaceutical industry research because of the demanding requirements for their design and contents. 

The isolation of DNA, which is suitable for digestion with restriction enzymes (which will be discussed in Section 6.1.7), is an essential requirement for both genetic engineering techniques as well as for the analysis of genetic makeup of industrial products. The isolation of DNA from various organisms requires specific protocols, and there are various companies that provide extraction kits. One vital element to remember (if one wishes to study with pure DNA preparations) is the contamination of the sample with other nucleic acids, namely RNA, as well as other macromolecules such as protein and polysaccharides. In order to avoid this, most extraction protocols include RNase for the removal of RNA, and proteinase K enzyme to remove protein contaminants [2]. 

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