From RNA to PCR template — Bulk RNA can be analyzed by PCR after reverse transcription into which nucleic acid form?

Difficulty: Easy

Correct Answer: c DNA

Explanation:


Introduction / Context:
Standard PCR requires DNA as a template. To analyze RNA transcripts, the RNA must first be converted into complementary DNA (cDNA) by reverse transcription, creating a DNA template suitable for amplification. This combined method is called reverse-transcription PCR (RT-PCR) or, in quantitative formats, RT-qPCR.



Given Data / Assumptions:

  • Input is RNA (e.g., total RNA or mRNA).
  • Reverse transcriptase synthesizes a DNA strand complementary to the RNA template.
  • Subsequent PCR uses DNA polymerase on this cDNA.


Concept / Approach:
Reverse transcription yields cDNA, a DNA copy of the RNA sequence. This cDNA is then amplified using PCR primers complementary to sequences of interest. tRNA and rRNA are specific RNA classes, not DNA forms; “m DNA” is not a recognized term. Therefore, the correct product enabling PCR from RNA is cDNA.



Step-by-Step Solution:

Isolate RNA from the sample (e.g., using spin columns).Use reverse transcriptase with primers (random hexamers, oligo-dT, or gene-specific) to generate cDNA.Perform PCR on cDNA with DNA polymerase to amplify targets.


Verification / Alternative check:
Successful RT-PCR shows amplicons even when no genomic DNA is present; minus-RT controls confirm the amplification derives from cDNA, not contaminating DNA.



Why Other Options Are Wrong:

  • tRNA / rRNA: RNA species, not DNA; cannot serve as DNA templates for standard PCR.
  • m DNA: not a standard term in molecular biology.


Common Pitfalls:
Forgetting DNase treatment of RNA to remove genomic DNA, which can otherwise confound RT-PCR results.



Final Answer:
c DNA

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