AdvancedSeq developed the SupreDye chemistry as a drop-in-ready alternative to BigDye chemistry for high-quality Sanger sequencing. Not only for the dyes, the PwrPOP polymers, ADS 5Xsequencing buffer, ADS 10X running buffer, and TruPure formamide we carry are also used interchangeably with the ABI Sanger sequencing counterparts, with comparable performance. Here, we will focus on the comparison between SupreDye v3.1 Cycle Sequencing Kit and BigDye v3.1 Terminator Cycle Sequencing Kit (for simplicity, we shorten the name below) and you can download our application notes for details. You can also download the performance comparison data between the two dyes using different types of templates.

The fluorescent signals generated from SupreDye chemistry decay slower over the run

One of the distinguishing features for SupreDye is the slower signal decay over the run compared to BigDye. This translates to better reading accuracy at the end of the readable region and would benefit users who are looking for high-quality long reading into the DNA sequence of interest. From both internal and external data, we see at least 50 bases can be read more accurately at the end.

Read-through of some high-GC content or secondary structure sequences

Unlike BigDye, the standard SupreDye cycle sequencing kit can read through some of the difficult templates we tested, such as templates with high-GC or secondary structure. For these templates, there is no signal drop through the difficult sequencing regions when the standard SupreDye kit, not the dGTP kit, is used. On the other hand, the dGTP kit is necessary to read through for BigDye. Both our internal and customer data have demonstrated this observation. Of course, the standard SupreDye kit can’t substitute for the dGTP kit when the template is much more difficult, such as templates containing long hairpin structure.

SupreDye dGTP kit generates much less compression for difficult templates

When SupreDye dGTP kit is used for a much more difficult template, we did not see compression issues. This is not true, and often reported, for the BigDye dGTP kit. We have found that the compression leads to missed reading of 15 bases over a long hairpin structure when the BigDye dGTP kit is used alone, although many suggest mixing of the standard and dGTP BigDye premix for improved compression. The fact that there is no need for mixing of the standard premix with the dGTP premix for SupreDye, helps save hands-on time.

If you would like to request a free sample of SupreDye v1.1 or v3.1 Cycle Sequencing Kit, you can contact us.