Planning a sequencing experiment? I’m sure you have a lot of questions like… Which is the right sequencing method for your application? Sanger or next generation sequencing? Let’s take a look at both in this episode of Seq It Out.
Before the 1990s, sequencing was done the “old fashioned” way; using gels and manually calling bases. Then, Sanger Sequencing by Capillary Electrophoresis was introduced. This let labs sequence long DNA fragments with automated base calls. As the technology evolved, Next Generation Sequencing, or NGS, transformed the way that sequencing was performed.
Let’s take a look at our lab book.
NGS is similar to Sanger sequencing in that you sequence DNA fragments. But in NGS, it’s massively parallel. This allows millions of fragments to be sequenced in a single run versus sanger sequencing which only produces one forward and reverse read. Researchers today have many choices when deciding which sequencing technology to use for their clinical research. Sanger sequencing with 99.99% accuracy is the “gold standard” for clinical research sequencing. However, newer NGS technologies are also becoming common in clinical research labs due to their higher throughput capabilities and lower costs per sample. But which one should you use? Sanger or NGS?
Here is what you have to consider. With its ability for long reads and gold-standard accuracy, Sanger sequencing is perfect for
- Sequencing single genes
- Sequencing 1-100 amplicon targets at the lowest cost
- Sequencing up to 96 samples at a time without barcoding
- Microbial Identification
- Fragment analysis, high throughput genotyping using, for example, SNaPshot
- Microsatellite or STR analysis
- NGS confirmation
Next generation sequencing offers fast turnaround time and takes only about 4 hours to complete a run. NGS is perfect for:
- Interrogating >100 genes at a time cost effectively
- Finding novel variants by expanding the number of targets sequenced in a single run.
- Sequencing samples that have low input amounts of starting material, using, for example, Ion AmpliSeq library preparation, which requires as little as 10 ng of input DNA
- Sequencing microbial genomes for pathogen subtyping to enable research of critical outbreak situations
With Applied Biosystems gold standard Sanger sequencing and simple, scalable Ion Torrent™ next-generation sequencing, Thermo Fisher Scientific has the right sequencing platform for every research application.
I hope this video was helpful on finding the right sequencing tool for your application, and I am sure you’ll have more questions. Submit your question at http://www.thermofisher.com/ask and subscribe to our channel to see more videos like this.
And remember, when in doubt, just Seq It Out