- Ultra deep sequencing providing highly sensitive detection levels without any bias
- Pooling of several samples possible
- Sequencing of PCR products without any time consuming cloning steps
- Supporting applications for diagnostic approaches
Amplicon sequencing is based on ultra deep sequencing of PCR products for analysing genetic variations. The long read lengths and data accuracy of Roche GS FLX makes the technology the gold standard for amplicon sequencing. As each amplicon molecule within a mixture of amplicons can be sequenced individually, the technology has the power to detect rare variants with detection limits of 0.5 % and even lower.
The amplicon variance analysis is ideally suited for highly variant samples, such as the hypervariable regions of antibodies, and samples with low variance, such as population samples for SNP detection.
Typical projects are:
- Diversity screenings within populations of individuals
- Identification of rare mutations associated with diseases (SNP detection)
- Metagenome analysis (e.g. 16S or 18S rDNA analysis)
- Study of methylation patterns
For amplicon sequencing, PCR products have to be linked with certain adaptors to generate appropriate libraries for next generation sequencing.
Amplifying your sequences of interest with a set of fusion primers is the method of choice.
A fusion primer consists of a target specific sequence at the 3' end and a fixed sequence at the 5’end for the sequencing approach (Primer A or Primer B). For sequencing several samples in parallel, you may pool several barcoded samples. Barcoding of amplicon samples is done by insertion of multiplex identifier tags (MIDs) into fusion primers (see figure).
Depending on your individual objective, we will assist you in designing the appropriate amplicon sequencing experiment for your research.
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