Reverse transcription polymerase chain reaction (RT-PCR) is a popular molecular method of identifying and analyzing RNA. It is usually used in gene expression research, viral diagnostics, and clinical diagnostics. A One-Step RT-PCR Kit streamlines this by doing both reverse transcription and PCR amplification in a single tube, thus saving on handling steps and minimizing contamination risk.
- Reverse Transcription: The conversion of RNA to cDNA
RT-PCR is first performed by transforming RNA into complementary DNA (cDNA). Reverse transcriptase is an enzyme that produces cDNA with the RNA template. This is necessary since PCR amplification is only accurate on DNA. Reverse transcription is accurate such that the original RNA sequence is faithfully represented in the resultant amplified product.
- PCR Amplification and Result Analysis
After making cDNA, the cycle of Repeat amplification with the application of DNA polymerase and primers is applied. The target DNA is doubled in every cycle, making it possible to detect the smallest quantities of RNA. The resultant amplified products are usually on agarose gels, using DNA ladders as size markers in order to verify the assumed size of the fragment and to interpret the results accurately.