Is There a Cost-effective Alternative to BigDye®?
Sanger Sequencing, the Gold standard for DNA Sequencing
Sanger sequencing has been around for 40 years. As a gold standard for DNA sequencing, Sanger sequencing provides 99.9% accuracy and has been widely used for multiple applications such as mutation identification and confirmation, de novo sequencing and resequencing. Because of its cost-effectiveness and ability to detect mutations in short DNA fragments in small sample settings, multiplex next-generation sequencing still can’t replace, but would in fact, need Sanger sequencing.
Chemistry of Dye terminator and Sanger Sequencing
The Sanger sequencing chemistry uses a small fraction of fluorescently labeled dideoxynucleotides (ddNTPS) mixed with dNTPs, the DNA building blocks to terminate the chain elongation by DNA polymerase. As a result, a cycle sequencing reaction with a template, a primer, and DNA polymerase produces a collection of extension products with only one nucleotide difference, with all of them fluorescently end-labeled. The extension products are then separated by capillary electrophoresis and fluorescent signals are detected by a genetic analyzer or sequencer. The final DNA sequence is composed through accurate basecalling on the software.
Cost-effective SupreDye Chemistry
Even with the decrease in cost and drop in price for Sanger sequencing, it is important to have high-quality, cost-effective dye chemistry alternatives. SupreDye chemistry works well with all different types of templates including ones with secondary structures or GC-rich content and produces high signal intensity and good signal uniformity through the whole long read length (see the SupreDye sequencing raw data below). We offer SupreDye Cycle Sequencing Kits for different templates and they can be used interchangeably with your standard dyes without changing the protocols.
Want to know more about SupreDye? Download our application note and contact us.