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    Polymerase chain reaction

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    Mary set up a similar PCR reaction. The Tm of the forward and reverse primers she uses is 55oC. The conditions that were optimized by her instructor were as follows:

    Cycle
    Temperature (oC)
    Time

    1X
    94o
    3 minutes

    30X
    94o
    30 seconds
    Denaturation
    50o
    30 seconds
    Annealing
    72o
    60 seconds
    Extension

    Mary sets up the PCR, but decides to see if she can get better results than her instructor, and makes the following changes in separate PCR runs.

    Describe the effect that each of her changes would have on the amount and purity of her PCR product. (Briefly explain).

    A.She changes the annealing temperature to 60oC

    B.She increases the number of cycles to 31

    C.She changes the magnesium concentration from 1.0 mM to 2.0 mM

    D.She adds non-complementary restriction enzyme sequences to the 3' ends of the primers

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    https://brainmass.com/biology/sequencing/polymerase-chain-reaction-152529

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    Mary set up a similar PCR reaction. The Tm of the forward and reverse primers she uses is 55oC. The conditions that were optimized by her instructor were as follows:

    Cycle
    Temperature (oC)
    Time

    1X
    94o
    3 minutes

    30X
    94o
    30 seconds
    Denaturation
    50o
    30 seconds
    Annealing
    72o
    60 seconds
    Extension

    Mary sets up the PCR, but decides to see if she can get better results than her instructor, and makes the following changes in separate PCR runs.

    Describe the effect that each of her changes would have on the amount and purity of her PCR product. (Briefly explain).

    A.She changes the annealing temperature to ...

    Solution Summary

    Answers to several questions regarding PCR are provided.

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