PCR Crash Course Part II

If PCR can’t be used to detect RNA, and coronaviruses are RNA viruses; there is no DNA and PCR does not replicate RNA, how can we detefct SARS-Cov-2?

1. use qPCR to detect DNA

2. we know of a method of creating RNA from DNA

What is RT-qPCR?

• is reverse transcriptase-qPCR

• Reverse transcription is an enzyme-mediated process by which complementary DNA (cDNA) molecules are created from RNA templates.

• After reverse transcription is completed, qPCR can continue as explained previously using cDNA as the template.

Performing RT-qPCR

1. sample (template)

2. Luna Enzyme (reverse transcriptase)

3. Primer Set

4. Probe Set

5. Luna Buffer (DNA polymerase)

Performing qPCR-Sample Concentration

Great sample concentration=?

Earlier detection of amplification

LoD (LoD)

The lowest concentration of target DNA that can be detected during amplification

In Lab Experiment

Synthetic Human DNA (P1)

Synthetic SARS-COV-2 RNA (N1)