Applied Biosystems SOLiD sequencing technology
SOLiD sequencing (Sequencing by Oligonuceotide Ligation and Detection) involves the generation of clonal bead populations via emulsion PCR. The major difference between SOLiD sequencing and other high-throughput sequencing platforms is its unique sequencing by ligation instead of sequencing by synthesis chemistry. Sequential ligations of fluorescently labeled probes detect every combination of two adjacent bases. Each base in the sequence is interrogated twice, and this dual interrogation contributes to increased base accuracy.
Step 1:
A fragment or mate-paired library is created, and used to create a clonal bead population.
Step 2:
Emulsion PCR is then carried out, and the PCR products are then covalently attached to a glass slide. A key advantage is the ability to accommodate high bead density per slide, resulting in a higher level of throughput.
Step 3:
Sequencing by ligation is carried out. Fluorescently-labelled dibase primers compete for ligation to the sequencing primer. Every 1st and 2nd base are interrogated in each ligation step, achieving a high level of specificity.
Step 4:
After a series of ligation cycles, the template is reset with a complementary primer at the n-1 position for a second round of ligation cycles. Multiple rounds of primer reset, followed by ligation cycles, ensure that virtually every base is interrogated at least twice in independent ligation reactions. This provides the SOLiD system with its unparalleled accuracy.
Specifications:
Read Lengths: Up to 50bp
Reads per run / Throughput: Fragment library: ~500M
System Accuracy: Greater than 99.94% accuracy due to 2 base encoding
Quality Score : >80% of bases at >QV30
Data Format: SOLiD native, SAM
Information taken from https://products.appliedbiosystems.com/ab/en/US/adirect/ab?cmd=catNavigate2&catID=604416&tab=TechSpec, accurate as of June 2010.
Applications:
- de novo sequencing and resequencing
- Chromatin immunopreciptation (ChIP)
- Methylation analysis
- Whole transcriptome analysis
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