The NGS amplicon sequencing is a powerful tool for biomarker research and diagnostic assay development. Avance Biosciences offers three different amplicon sequencing approches with a focus on scalability, speed, and cost. State-of-the-art Illumina MiSeq platform is used. GLP/GMP-compliant service is available.
One-step PCR Method
One-step PCR method requires a pair of custom designed PCR primers that include target-specific sequences, indices, and NGS adaptors. This approach is specially useful for developing diagnostic assays where routine testing of same target sequence is performed.
Two-step PCR Method
This method include two PCR steps. The first PCR uses primer pairs containing target-specific PCR sequences and sequence tags used as primer sites for a second PCR. The second PCR incorporates sequencing indices and library adaptors into the final amplicons ready for sequencing. If you are interested in analyzing a few (1-3) amplicons on a large number of samples, this method the most flexibility and fast turn-around time.
When large number of amplicons are sequenced, the extension-ligation method is your choice. With our specially designed primer pairs, amplicons of interest are first pull-down as single-stranded fragments, which are then PCR amplified. This method allows hundreds of regions to be sequenced simultaneously while minimizing the bias of PCR.
For more information, please contact our technical staff.