Nurturing Life Sciences
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#Unit 3. Fundamental Processes
A common method to monitor synthesis of new DNA is Incorporation Assays.
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rNTPs are present at approximately 10-fold higher concentration in the cell, they are incorporated at a rate that is more than 1000-fold lower than dNTPs.
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The palm domain is composed of a b sheet and contains the primary elements of the catalytic site.
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DNA polymerase binds two divalent metal ions (typically Mg2+ orZn2+) that alter the chemical environment around the correctly base-paired dNTP and the 3’-OH of the primer
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One metal ion generate a 3’O- that is primed for the nucleophilic attack of the a-phosphate of the incoming dNTP.
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The second metal ion coordinates the negative charges of the b-phosphate and g-phosphate of the dNTP and stabilizes the pyrophosphate produced by joining the primer and the incoming nucleotide