What is the Difference Between Conventional Nested and Real-time PCR Assays?
🆚 Go to Comparative Table 🆚The main differences between conventional nested and real-time PCR assays are as follows:
Conventional PCR (C-PCR):
- Developed to amplify specific fragments of DNA.
- Single amplification reaction with a single set of primers.
- Less sensitive compared to nested and real-time PCR.
Nested PCR (N-PCR):
- Developed to improve the sensitivity of conventional PCR.
- Requires two successive amplification reactions with two separate sets of primers.
- The first round of amplification is performed with the outer primers, and the product is then used as a template for the second round of amplification with the inner primers.
- More sensitive than conventional PCR but can give erroneous positive results due to DNA contamination.
Real-time PCR (Q-PCR):
- Developed to quantify the amplified PCR product.
- Able to provide results within 2 hours from genomic DNA extraction to data analysis.
- Highly sensitive and specific.
- Potential for automation and high-throughput analysis.
In summary, conventional PCR is the original technique for amplifying specific DNA fragments, while nested PCR is a variation that improves sensitivity by using two successive amplification reactions with two sets of primers. Real-time PCR, on the other hand, is a more advanced technique that allows for the quantification of amplified PCR products and offers higher sensitivity and specificity compared to conventional and nested PCR assays.
Comparative Table: Conventional Nested vs Real-time PCR Assays
Here is a table comparing the differences between conventional nested and real-time PCR assays:
Feature | Conventional Nested PCR | Real-Time PCR |
---|---|---|
Sensitivity | Lower | Higher |
Specificity | High | High |
Detection Method | gel electrophoresis | Fluorescence |
Quantification | Not quantitative | Quantitative |
Processing Time | Longer | Shorter |
Labor Intensity | More labor-intensive | Less labor-intensive |
Applications | Various genetic analyses | Various genetic analyses |
Both conventional nested and real-time PCR assays are modifications of conventional PCR and are used to amplify DNA samples for various genetic analyses. However, real-time PCR is a more sensitive and specific technique that allows for the quantification of amplified products, which is not possible with conventional nested PCR. Real-time PCR also has the advantage of shorter processing time and being less labor-intensive compared to conventional nested PCR.
- PCR vs Real-time PCR
- RT PCR vs QPCR
- Rapid vs PCR Test
- NAAT vs PCR
- Consensus PCR vs Pan PCR
- PCR vs DNA Sequencing
- Continuous Assay vs Stopped Assay
- Gene Cloning vs PCR
- LAMP vs PCR Test
- Microarray vs RNA Sequencing
- PCR vs DNA Replication
- Micronucleus vs Comet Assay
- ELISA vs Rapid Test
- Biochemical vs Cell Based Assays
- Microarray vs Next Generation Sequencing
- DNA vs RNA Probes
- Colorimetric vs Fluorometric Assay
- PCR Primers vs Sequencing Primers
- rDNA vs cDNA