RT-PCR involves all of the following except which step?

RT-PCR involves all of the following except which step?

• A. DNA isolation
• B. cDNA synthesis
• C. PCR amplification
• D. Gel electrophoresis

Answer: (A)

RT-PCR uses RNA as the starting template, not DNA. The workflow begins with isolating RNA from the sample, then reverse transcribing that RNA into complementary DNA (cDNA), and finally amplifying the cDNA by PCR. Detection of the amplified product can be done in real time or by post‑PCR methods such as gel electrophoresis, depending on the protocol. DNA isolation isn’t part of the standard RT-PCR process because you’re not starting from DNA or converting DNA; you’re working with RNA and converting it to cDNA first.

RT-PCR uses RNA as the starting template, not DNA. The workflow begins with isolating RNA from the sample, then reverse transcribing that RNA into complementary DNA (cDNA), and finally amplifying the cDNA by PCR. Detection of the amplified product can be done in real time or by post‑PCR methods such as gel electrophoresis, depending on the protocol. DNA isolation isn’t part of the standard RT-PCR process because you’re not starting from DNA or converting DNA; you’re working with RNA and converting it to cDNA first.