What is the Difference Between NGS and Sanger Sequencing?
🆚 Go to Comparative Table 🆚The main difference between Next-Generation Sequencing (NGS) and Sanger Sequencing lies in the sequencing volume and the number of DNA fragments that can be sequenced simultaneously. Here are the key differences between the two methods:
- Sequencing Volume: Sanger sequencing sequences a single DNA fragment at a time, while NGS is massively parallel, sequencing millions of fragments simultaneously per run.
- Throughput: NGS has a higher throughput, allowing for the sequencing of hundreds to thousands of genes at once, while Sanger sequencing is limited to a smaller number of samples.
- Discovery Power: NGS offers greater discovery power to detect novel or rare variants with deep sequencing.
- Sensitivity: NGS has a higher sensitivity compared to Sanger sequencing, which can only detect variants when they occur in about 15 to 20 percent of the sequences.
- Accuracy: Sanger sequencing is considered the "gold standard" for clinical research sequencing, with an accuracy of 99.99 percent. However, NGS methods are becoming more common in clinical research labs due to their higher throughput.
- Applications: Sanger sequencing is suitable for sequencing single genes, 1-100 amplicon targets, up to 96 samples at a time, microbial identification, and fragment analysis. NGS is more suitable for large-scale sequencing projects and detecting multiple variants across targeted regions.
In summary, Sanger sequencing is more suitable for smaller projects and validation of gene variants, while NGS is more suitable for large-scale sequencing projects and detecting multiple variants across targeted regions. Both methods have their strengths and limitations, and the choice between them depends on the specific requirements of the research or application.
Comparative Table: NGS vs Sanger Sequencing
Here is a table comparing the differences between Sanger sequencing and Next-Generation Sequencing (NGS):
| Feature | Sanger Sequencing | Next-Generation Sequencing (NGS) |
|---|---|---|
| Sequencing Volume | Sequences a single DNA fragment at a time | Massively parallel, sequencing millions of fragments simultaneously per run |
| Speed | Relatively slow | Much faster |
| Accuracy | Very accurate (99.99%) | Less accurate than Sanger sequencing |
| Fragment Length | Can only sequence relatively short DNA fragments | Can sequence much larger DNA fragments |
| Applications | Ideal for sequencing small regions of DNA with high accuracy | Suitable for large genomes or applications where speed and cost are important |
| Sensitivity | Lower sensitivity, detects variants when they occur in about 15 to 20 percent of the sequences | Higher sensitivity, better for detecting rare variants |
Sanger sequencing is a highly accurate method for sequencing small regions of DNA, but it is limited in speed and the length of fragments it can sequence. On the other hand, NGS is a faster and more cost-effective method that can sequence larger DNA fragments simultaneously, making it suitable for applications where speed and cost are important.
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