Sequencing by Oligonucleotide Ligation and Detection
Developed by Life Technologies and made available in 2006, the SOLiD 5500xl sequencer is the only next generation platform to sequence via primer ligation. This method of sequencing provides internal accuracy checks as each ligation is coded by two nucleotides. As a result, each nucleotide is sequenced twice and the overall accuracy of the sequencing data is 99.94% – one of the highest on the market.
The basic principle behind how SOLiD sequencing works is fluorophore detection. First, libraries undergo emulsion PCR to clonally amplify single library molecules onto beads. During sequencing, oligos are pushed through a flow cell and allowed to anneal to the library bead. Each sequencing oligo is tagged with one of four fluorophores. When an oligo finds and anneals to its complementary sequence on the template, an enzymatic reaction takes place that cleaves off the fluorophore. A camera then takes a picture of each color emission, the data is recorded in “color-space”, and the software converts the color data into base-space.
To become a member of the SOLiD community, create an account at www.abcommunities.lifetechnologies.com.
SOLiD Sequencing Applications
There are several supported applications for SOLiD sequencing. Life Technologies offers a series of Webinars for users to get information on their most popular applications.
Here are some application notes for some of the more widely-used SOLiD applications:
- Whole Genome Resequencing
- Small RNA Sequencing
- ChIP-Seq
- Whole Transcriptome applications such as Exome-Exome Junction Mapping and Fusion Transcript sequencing and Single cell whole transcriptome sequencing.