Aierward the sequence can be read o by going from top

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Unformatted text preview: anger Sequencing A G C Primer C G A C T A C T A C T ConNnue unNl all strands of DNA have undergone this reacNon. If you choose the reagents correctly then you should have all possible A ­terminated strands; resulNng in sequences of varying lengths. Sanger Sequencing Sanger Sequencing In the radioacNve gel, the longer DNA fragments move to the bopom and the shorter ones move to the top. Aierward the sequence can be read off by going from top to bopom. To recap… •  Sanger Sequencing: –  Run a PCR reacNon in the presence of a bunch of ddNTPs, with each different base pair dyed a different color. –  Measure the length and color of the resulNng fragments of DNA, and use that to work out the sequence. •  Requires a lot of space and Nme: you need a place to run the reacNon, and then you need a capillary tube or a gel to determine the length of the DNA. –  You could only run perhaps a hundred of these reacNons at any one Nme. –  There are 3 billion base pairs of DNA in the human genome, meaning about 6 million 500 ­base pair fragments of DNA. 40 Celera Sequencing (2001) •  •  •  •  •  300 ABI DNA sequencing plaXorms 50 producNon staff 20,000 square feet of wet lab space 1 million dollars / year for electrical service 10 million dollars in reagents Total cost of human genome: 2.7 Billion dollars Celera Sequen...
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