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What would the correct answer for this question be and why? And why would the other choices be wrong?

Question 85 Which of the following would most strongly reduce the strength of a real-time colony PCR? O A. 16S rRNA has minor SNPs between different species of bacteria. O B. Consistently running the PCR amplification for one O C. Different species of bacteria have different numbers O D. Different species of bacteria express 16S rRNA in extra cycle. of 16S rRNA gene copies. different levels.

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Answer . B) consistently running the PCR amplification for one extra cycle because adding one more cycle can reduce the strength of a real time PCR as the one more cycle leads to doubling amount of product which produced in previous cycle. If the added dNTP are less then it will reduce the strength of real time PCR.

16S rRNA has minor SNPs between different species of bacteria is false because in PCR only primer are needed for amplification of 16S rRNA. These SNPs are present within sequence so it will not reduce strength.

Also primer used for real time PCR is universal so different species of bacteria either express 16S rRNA in different level or different number of 16S rRNA gene copies will not affect strength of real time PCR

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