What is the Difference Between RT PCR and QPCR?
🆚 Go to Comparative Table 🆚The main differences between RT-PCR and qPCR involve the purpose, the technology used, and the workflow. Here are the key distinctions:
- Purpose: PCR (polymerase chain reaction) amplifies a specific DNA sequence, while RT-PCR (reverse transcription PCR) is used to detect and measure the amount of RNA by converting it into complementary DNA (cDNA) first. qPCR (quantitative PCR), on the other hand, is an advanced method used to amplify and quantify the amount of DNA or RNA in real-time.
- Technology: PCR typically uses endpoint reactions, while qPCR and RT-qPCR (reverse transcription quantitative real-time PCR) use the kinetics of the rate of product synthesis during the PCR reaction to quantitate the amount of template present. qPCR can be further divided into two approaches: dye-based qPCR, which measures the fluorescence of a dye that binds to double-stranded DNA, and probe-based qPCR, which uses fluorescence-labeled DNA probes.
- Workflow: In RT-PCR, RNA is isolated, and cDNA is generated via reverse transcription. PCR is then carried out to amplify areas of interest. In qPCR, DNA is isolated and amplified, with amplification quantitated using a fluorescent probe. RT-qPCR combines RT-PCR with qPCR, enabling the measurement of RNA levels and allowing for rapid detection of changes in gene expression.
In summary, PCR is a basic technique for amplifying a specific DNA sequence, while RT-PCR is used for detecting and measuring RNA levels by converting it into cDNA. qPCR is an advanced method for real-time amplification and quantification of DNA or RNA, and RT-qPCR combines the reverse transcription step with qPCR for more accurate and rapid gene expression analysis.
Comparative Table: RT PCR vs QPCR
Here is a table comparing the differences between RT-PCR and qPCR:
| Feature | RT-PCR | qPCR |
|---|---|---|
| Definition | Reverse Transcription PCR, used to detect and measure RNA. | Quantitative PCR, used to amplify and quantify DNA. |
| Template | RNA | DNA |
| Process | Involves reverse transcription and PCR. | Involves PCR with fluorescence to measure amplification product. |
| Quantitation | Measures RNA levels. | Measures DNA levels, can determine copy number of DNA sequence. |
| Applications | Gene expression analysis, RNAi validation, microarray validation, etc. | Pathogen detection, genetic testing, disease research, etc. |
| Steps | Reverse transcription, PCR. | PCR with fluorescence measurement. |
Please note that RT-PCR stands for Reverse Transcription PCR, while qPCR stands for Quantitative PCR. Both techniques are used in molecular biology laboratories for various applications, such as gene expression analysis, pathogen detection, and genetic testing.
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