184x Filetype PPTX File size 2.22 MB Source: w3.biosci.utexas.edu
Overview Southern-blot DNA Digestion Based hybridization COBRA MSP Bisulfite Sequencing Bisulfite Based Real-time MSP MethyLight MassARRAY Pyrosequencing Southern Blot Hybridization 1978 -One of the older techniques for sequencing (1978), not as widely used any more due to the development of PCR and other cheap/fast sequencing techniques, not outdated but just not as widely employed Process: • Step 1 use restriction enzymes to cut DNA • Step 2 electrophoresis to separate DNA fragments by size • Step 3 transfer DNA fragments from gel to a membrane • Step 4 hybridize radiolabeled probe to membrane • Step 5 sample is washed to remove extraneous radio probes Southern Blot Hybridization 1978 Advantages: • Cheap • simple • 1 probe can detect multiple similar sequences • well suited for analyzing long stretches of DNA Disadvantages • Requires a large amount of DNA • Time consuming • labor intensive • limited sensitivity Interesting probe gif http://www.slic2.wsu.edu:82/hurlbert/micro101/images/101SouthernBlot10.gif
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