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09Ch18&19ex3 - Lauren Taranow lmt827 8 am...

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Lauren Taranow lmt827 8 am Discussion Questions and Problems for Chapters 18 and 19 Material From Brooker - none Additional Question A2. Explain how to do each of the following procedures, what it accomplishes (why you would do it), and how it works. a. automated sequencing Automated sequencing, similar to replication, is a way to determine the sequenceof a cloned gene, and requires a DNA polymerase and a primer. DNA is denatured by adding heat, and DNA polymerase adds a high concentration of dNTPs, as well as incorporating a low concentration of fluorescently-marked ddNTPs that terminate the nucleotide chain. The chain-terminating ddNTPs are randomly encorporated at every position along the insert. Frgments are separated in a PAGE gel, and as fragments run off the end of the gel, they are hit by a laser to excite the fluorophores attached to the ddNTP. The fluorescence is read by a detector, which reports the colors in the order they are emitted to tell us the unknown sequence of nucleotides.
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