PCR variants

What are the three steps in PCR with each corresponding temperature?

• Denaturation, Annealing, Elongation at 90°C, 40°C, 60°C

• Denaturation, Annealing, Elongation at 37°C, 55°C, 72°C

• Denaturation, Annealing, Elongation at 50°C, 60°C, 70°C

• Denaturation, Annealing, Elongation at 95°C, 50°C, 72°C

What is the purpose of Reverse Transcription in RT-PCR?

• To convert RNA to cDNA

• To detect RNA using RNA polymerase

• To amplify RNA using DNA polymerase

• To convert DNA to RNA

What is the main advantage of Multiplex PCR over Traditional PCR?

• Multiplex PCR can detect multiple targets in a single reaction

• Multiplex PCR consumes more time and effort

• Multiplex PCR is more prone to contamination

• Multiplex PCR is less sensitive

What is the purpose of Nested PCR?

• To reduce sensitivity and specificity

• To eliminate the need for gel electrophoresis

• To improve sensitivity and specificity

• To decrease the number of cycles needed

What is the main advantage of qPCR over Traditional PCR?

• qPCR can determine starting DNA concentration with accuracy

• qPCR requires postamplification manipulation

• qPCR is more cost-effective than Traditional PCR

• qPCR is less accurate in determining DNA concentration

What is the purpose of the Baseline in real-time PCR?

• To indicate the end of the amplification

• To measure the starting DNA concentration

• To show the level of signal during initial cycles

• To determine the cycle threshold

What does the Cycle Threshold (CT) value represent in qPCR?

• The cycle at which the amplification plot crosses the threshold

• The total number of cycles needed for amplification

• The cycle at which the amplification reaches the plateau phase

• The cycle at which the baseline signal is detected

What are the two common methods for detecting PCR products in Real-Time PCR?

• Non-specific fluorescent dyes and RNA probes

• Non-specific fluorescent dyes and DNA probes

• Specific fluorescent dyes and DNA probes

• Specific fluorescent dyes and RNA probes

What does CT value of 10 indicate in a Real Time PCR test?

• Positive result

• Negative result

• High concentration of virus

• Low concentration of virus

Which is the advantage of using EvaGreen over SYBR Green in PCR?

• Less inhibitory to PCR

• Causes mispriming

• Inhibits PCR

• Unsuitable for high resolution melt curve analysis