What are the pros and cons of RT-PCR?

Answer 1

There are several advantages of using real-time PCR over other methods. Real-time PCR assays are thousand folds more sensitive than RNase protection assays or dot blot hybridization. It allows you to quite precisely calculate and compare of the amount of template in each cycle, instead of determining the amount of product at the end of the reaction.

Quantitative RT-PCR is commonly used for clinical applications. For example, you could use this method to quantify the amount of HIV RNA particles per ml of blood plasma in a patient who is undergoing treatment with antiviral drugs to see if it's working or not.

The main problem with real-time PCR is that it requires specialised thermal cyclers (PCR machine) with fluorescence monitors and its reagents are quite expensive.

Answer 2

Pros: RT-PCR is highly sensitive and specific, allowing for detection and quantification of RNA targets. It is widely used for diagnosing Infectious Diseases, monitoring gene expression, and detecting genetic mutations.

Cons: RT-PCR requires specialized equipment and trained personnel, making it relatively costly and less accessible in resource-limited settings. There is also a potential for contamination during the amplification process, leading to false-positive results.