Between standard PCR and reverse transcription PCR techniques, which one would you employ to amplify a desired gene from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Explain why
Added by Michael B.
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SARS-CoV-2 is an RNA virus, meaning its genetic material is single-stranded RNA, not DNA. Show more…
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QUESTION 1 [30] 1.1 Compare the theory and practice behind conventional polymerase chain reaction (PCR), real-time PCR, and DNA sequencing. In your answer, mention all the enzymes and reagents used in these assays with their functions (20) 1.2 Explain the use of the enzyme reverse transcriptase in a SARS-CoV-2 molecular diagnostic assay. (10)
Sri K.
The DNA polymerase chain reaction (PCR) is a highly used biochemical method to amplify minute quantities of DNA as much as 1,000,000-fold. It has found many applications, including the diagnosis of COVID-19 patients after infection with the SARS coronavirus. Because PCR amplifies DNA only, to test for the presence of COVID-19 (which is an RNA virus), reverse transcriptase must be used first to convert the viral RNA into DNA. As shown here, PCR involves the use of the following components: 1) two oligonucleotides (small red arrow and small green arrow) that are complementary to different positions within the DNA (COVID-19 sequences) to be amplified, 2) DNA polymerase, and 3) successive heating and cooling steps that denature and then anneal DNA strands. Based on your knowledge of the complementarity of DNA and the synthesis of DNA by using DNA polymerase, explain how PCR works.
Madhur L.
Answer the following question: Which of the following statements about qPCR is incorrect? a. RT-qPCR means reverse transcription-quantitative PCR, which is a method researchers developed to transcribe RNA to DNA first and then report the progress of PCR in real time. b. For COVID-19 detection, qPCR instead of the conventional PCR method is chosen because the former allows for multiplex detection and the signals are generated by equipment in a massive manner. c. The higher the initial amount of sample, the higher deltaRn value will be obtained. d. qPCR is the gold standard chosen by the CDC for COVID-19 detection.
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