A microbiologist is working with PCR and wishes to quantify the PCR by monitoring the amplification process while the PCR is ongoing. Which of the following types of PCR should he do?
Real time polymerase chain reaction (PCR) can be used to quantify the PCR by monitoring the amplification process while the PCR is ongoing. Types of PCR: Reverse transcriptase PCR: The enzyme reverse transcriptase () is used to synthesise a complementary copy of DNA (cDNA) from RNA and this cDNA is then used a template for PCR. Real-time PCR: This helps quantify the PCR and gives an estimate of the pathogen DNA load in the specimen. It uses fluoroscent dyes or specific DNA probes. Multiplex PCR: Primers to more than one pathogen are used simultaneously and hence multiple products are obtained. Nested PCR: It involves the sequential use of two primer sets. First primer set is used for amplification and the amplicon obtained is then used as a target sequence for second amplification. This ensures specificity. Reference Ananthanarayan and Paniker&;s Textbook of Microbiology Tenth edition
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