Lab Report #4mol 215

Lab Report #4mol 215 - Lab Report #4 Isolation of Your Own...

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Lab Report #4 Isolation of Your Own DNA for Amplification By PCR 1. In PCR, the temperatures exceed the maximum levels that human or E coli DNA polymerases can handle. At temperatures as high as 95 o C, the enzymatic half-life of Human polymerase is much too small to be functional for the duration of the PCR process. The Taq polymerase is a thermostable polymerase that remains functional for longer periods of time at the high temperatures PCR requires during denaturing. (It has a half-life of between 40 and 90 minutes at 95 o C) Taq polymerase has a temperature optimum of about 72 o C.Human DNA polymerase works most efficiently at around body temperature or 37 o C. This is significantly lower than the temperatures used for denaturing the DNA (95 o C). DNA tends to “melt” or denature somewhere between 75 o C and 98 o C depending upon the organism. The human DNA polymerase would extend the new DNA strand and then be rapidly denatured during the Denaturing process of the next cycle. Human polymerase would have to be added every cycle (for 30 or more cycles) in
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This note was uploaded on 09/20/2008 for the course MOL 215 taught by Professor Edwardc.cox,philipg.felton during the Fall '06 term at Princeton.

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Lab Report #4mol 215 - Lab Report #4 Isolation of Your Own...

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