Bacteria genomic DNa

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Catalog | Cart | Log In Corporate Products Resources Applications | Features | LabFacts | FAQs | Cartoons | Site Help Busy? Don't miss an article! Get Pub Hub e-mail updates or automatic updates (via RSS feed). Focus: DNA Purification Isolation of Genomic DNA from Bacterial Cells using the Wizard ® SV Genomic DNA Purification System These experiments describe isolation of genomic DNA from a variety of bacterial samples using the Wizard ® SV Genomic DNA Purification System ( Cat.# A2360, A2361 ) By Ryan McDonald, M.Sc., and Olga Hill, MLT. Molecular Diagnostics, Provincial Laboratory, Saskatchewan Health, Regina, SK, Canada
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Introduction With the emergence of antimicrobial-resistant organisms and the increased prevalence of food-borne human pathogens, surveillance of these bacteria has become an important interest to public health. For example, methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of nosocomial infections in health care facilities in many countries. Enteric bacteria such as Salmonella and Escherichia coli pose a threat to public health as causes of food-associated outbreaks. Molecular-based microbial subtyping methods have been refined over the past few years to aid in epidemiological investigation of outbreaks. One method, fluorescence-based Amplified Fragment Length Polymorphism (fbAFLP) DNA fingerprinting, has become a valuable technique for characterizing bacterial strains (1– 3). For a clinical laboratory to be able to respond to increased numbers of outbreaks and isolates, methods such as fbAFLP must be adapted to handle high volumes by increasing throughput while decreasing turnaround-time. fbAFLP requires a good yield of high quality genomic DNA from each isolate. Conventional methods are labour-intensive and time- consuming, often requiring intricate manipulations and the use of hazardous organic chemicals. This results in a bottleneck in the fbAFLP procedure that limits the overall throughput and turnaround time for communication of results to the appropriate health care personnel. Introduction of an alternative DNA purification system must be considered to address these concerns. Here we describe an application of the Wizard ® SV Genomic DNA Purification System for the purification of genomic DNA from Gram-positive and Gram-negative human pathogens including Staphylococcus aureus , Staphylococcus epidermidis , Enterococcus faecalis , Listeria monocytogenes , Salmonella Typhimurium, Salmonella Heidelberg and Escherichia coli O157:H7. The genomic DNA is isolated from overnight broth cultures and yields high quality DNA that serves as a good template for restriction/amplification analyses such as fbAFLP. Methods and Results
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This note was uploaded on 10/03/2011 for the course CHEM 113A taught by Professor Professornotknown during the Spring '09 term at San Jose State.

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