Bi1X_2009_Week3_prelab_PCR

Bi1X_2009_Week3_prel - Bi1x Spring 2009 Week 3 Prelab Understanding PCR One of the key steps in our experiment is amplifying the environmental

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Bi1x – Spring 2009 Week 3 Prelab: Understanding PCR One of the key steps in our experiment is amplifying the environmental genomic DNA that we have extracted from the Caltech ponds. We will achieve this amplification using the ubiquitous process of Polymerase Chain Reaction (PCR). As shown in Fig. 1 below, through a repetitive series of temperature cycles, DNA polymerase acts as a molecular Xerox machine copying molecules again and again. The goal of this Prelab is to get you acquainted with the concept of PCR. Problem 1 A. Templates, amplicons and anchored products in a PCR While it is true that the total number of molecules in a PCR reaction increases exponentially as a function of the cycle number, closer inspection reveals that this population is actually the sum of three distinct populations at any given moment in the PCR tube: there is the "original template", the "amplicons" and the "anchored products", all of which are targets for further amplification. The original template is simply the original molecule(s) from which we are trying to amplify a certain sequence. The amplicon is the double stranded DNA that stretches from the sequence of the forward primer to the sequence of the reverse primer; this is the desired end product. The
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Bi1X_2009_Week3_prel - Bi1x Spring 2009 Week 3 Prelab Understanding PCR One of the key steps in our experiment is amplifying the environmental

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