Since we’ll be talking a lot about sequencing and sequencing results in the next few weeks, we’d like to start by telling you about the technology we used.
Generally speaking if you want to sequence something today there are two approaches: Sanger sequencing or next-generation sequencing (NGS) methods. NGS methods are the go-to methods if you want to sequence a complete genome (as we did with LilBUB), because the cost to sequence a single base (the building blocks of DNA) is much less and you can work at high-throughput (sequence a lot of DNA at the same time). Having said that it’s still a couple of thousand dollars per genome, so Sanger sequencing may be a better choice if you’re only interested in sequencing a small segment of DNA. Previously, we used Sanger sequencing to find the mutation that’s responsible for BUB’s polydactyly.
This time, to sequence the whole genome of Lil BUB we used NGS, though. More specifically, we used Illumina sequencing. There are some excellent videos and resources describing the technologies behind NGS, so rather than adding to that, here a selection, allowing you to browse:
1. A 2 minute animation by DrDoubleB. If you don’t want too much technical detail this is the one to watch.
2. A really good overview and comparison of different NGS methods by abm. It’s pretty detailed and technical, but they also have a great website with lots of information and a graphical description you can browse.
3. And finally, here’s an online course by Illumina (the link leads you to the main training site, select Sequencing: Illumina Technology) if you want a step-by-step explanation of the specific method we used. Below you can also find a video, which is a pretty accurate representation of what we did, although our sample preparation method was a bit different from the one described.
In our next post we’ll describe why we selected the Illumina technology.