Systems Microbiolo23

Systems Microbiolo23 - the genome o Both approaches have...

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Systems Microbiology o Later people realized that you can use fluorescent labels instead of radiolabels. ° This meant that you didn’t have to deal with radioactivity ° It also meant that you could run them all in one lane. ° Instead of a slab gel, people use a thin tube, with a fluorescence detector automatically reading the wavelengths as they come out the other end. ° This method is fast and accurate o Graph: the progress of the Human Genome Project o Shotgun genome sequencing – what about the gaps? ° Doing a complete sequence for a bacterial genome is still expensive – around 10 or 20 thousand dollars, but it’s relatively straightforward. o Small insert cloning/sequencing o There are automated plate pickers now o Some people thought that shotgun sequencing would work (the Celera group used this) but others thought that it wouldn’t (the public consortium used a different method) o BAC sequencing – larger segments, you know where they lie on
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Unformatted text preview: the genome o Both approaches have advantages: shotgun sequencing is much quicker, but BAC sequencing closes the gaps – the best method is to use some of both. o 454 Sequencing – Commercial, massively parallel DNA sequencing technology: ° Run PCR with beads, each of which attaches to on strand of DNA ° Emulsion PCR ° Lots of DNA comes from an individual strand. ° You pour the reaction over a chip with tiny wells that each fit one bead. ° 1.5 million reactions all at once ° Thus the factory approach to sequencing is no longer necessary – it can all be done in little machines like this. ° It’s still pretty expensive, but it’s getting closer, and it’s changing things rapidly. o The evolution of sequencing is moving towards nano-methods, and pretty soon there will probably be sequencing technologies in hospitals....
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