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ses2_ln - MIT OpenCourseWare http/ocw.mit.edu 7.344...

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MIT OpenCourseWare http://ocw.mit.edu 7.344 Directed Evolution: Engineering Biocatalysts Spring 2008 For information about citing these materials or our Terms of Use, visit: http://ocw.mit.edu/terms .
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Session 2 Lecture Notes 1. Ask again if everyone understands PCR. Some terms to discuss: Hot start and prolonged extension. Why would one use these? Hot start inhibits polymerase activity during sample preparation thereby eliminating non- specific amplification. Prolonged extension allows for final extension to be completed. (especially if large or GC rich because Tm and annealing temp is higher.) 2. How is the gene of interest selected? Vector contains large deletions in the ribozyme gene. 3. What is the method and how is it done? Authors vary the amounts of individual bases 4. What are the results of this paper? Look at Table 1 and Figure 3. Method identified for PCR that gives an overall mutation rate of 0.66% with no strong mutational bias. What is mutational bias? Is that obvious in this procedure? (Less diversity).
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