RT-PCR · Pathology

When performing RT-PCR, why is it often necessary to treat the RNA sample with DNase I prior to reverse transcription?

To stabilize the RNA molecules
To remove genomic DNA from the RNA preparation
To act as a cofactor for the reverse transcriptase
To facilitate the binding of random hexamers

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Correct answer: To remove genomic DNA from the RNA preparation

Genomic DNA contamination can be co-amplified during the PCR step, leading to inaccurate quantification or false positives. DNase I digests DNA without harming the RNA template, ensuring that the final signal originates solely from the RNA.

Difficulty: Medium Question 5 of 20

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When performing RT-PCR, why is it often necessary to treat the RNA sample with DNase I prior to reverse transcription?

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