explain why a quantitative pcr analysis cannot determine the size of the initial template the sequence in a dna sample.

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01-12-2022
Biology

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explain why a quantitative pcr analysis cannot determine the size of the initial template the sequence in a dna sample.

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Paruljain Paruljain · Answer 1 · 2022-12-05T12:20:47-05:00

Quantitative PCR analysis cannot determine the size of the initial template sequence in a DNA sample just because it measures the amount of particular sample present.

Building on the fundamental PCR technology, quantitative PCR (also known as quantitative real-time PCR, or qPCR) detection enables scientists to determine how much starting material is present in a sample. qPCR has a much greater dynamic range of analysis than traditional, end-point PCR since the products are detected as the reaction progresses; from one copy to approximately 1011 copies are recognized in a single run. Using a closed-tube format for quantitative real-time PCR and the subsequent amplicon identification, which does not require post-PCR processing such as gel electrophoresis, dramatically reduces the possibility of cross contamination. In order to determine indirectly how much nucleic acid is present during each cycle of amplification, qPCR uses a fluorescent reporter dye. During the reaction's repeating stages, the number of exponentially increasing PCR product molecules (amplicons) generated is directly proportional to the growth in fluorescence signal. Reporter molecules can be classified into one of three groups: additional dye-conjugated oligonucleotides known as probes, dyes attached to primers.

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