Targeted resequencing studies require the ability to sequence large numbers of samples across specific regions of interest. Targeted resequencing allows researchers to obtain statistically significant data sets for their regions of interest from a large number of samples while avoiding the expense and time required for whole genome sequencing. The challenge of a successful targeted resequencing project is combining the massive throughput of next generation sequencers—which are traditionally optimized for sequencing the entire genome of a single sample at a time—with the experimental needs of sequencing many samples across specific regions. The Access Array System enables a new level of targeted resequencing by combining PCR-based target enrichment capability with amplicon tagging. Now, users can take full advantage of their next-generation platform and sequence hundreds of samples per run.
The Access Array Integrated Fluidic Circuit (IFC) facilitates
parallel amplification of 48 unique samples, in effect preparing 48
sequencing libraries, in just a few hours. Every reaction combines both
an amplicon tagging and a barcoding (identification) step that enables
all 48 samples to be multiplexed at the sequencing step. This powerful
chemistry simplifies the up-front preparation and maximizes the
utility of today's targeted genome sequencing and next-generation sequencers.
The Fluidigm Access Array System can be used with any PCR-based
sample preparation method and with the reagents and primers of your
choice. The system includes Access Array IFCs, two IFC Controller AXs, and the Stand-Alone Thermal Cycler. These components deliver sample throughput and coverage scalable with laboratory
|Target Enrichment of Genomic DNA Regions|
enrichment refers to the ability to select a specific region of
interest prior to sequencing. For example, if you were interested in
examining 20 specific genes from a large cohort of individuals it would
be both wasteful and prohibitively expensive to sample the entire genome
of each individual. Instead, target enrichment strategies allow you to select regions for amplification from each individual and thus only sequence the specific area of interest.
|Sample Barcoding for Multiplexed Sequencing|
of the largest challenges facing next-generation sequencing operators
today is how to use the massive amounts of throughput enabled by the
new crop of sequencing equipment. While all of the systems in use today allow
massive amounts of data to be generated on a per-sample basis, they lack
a simple and reliable method for running multiple samples per sequencing run.
Barcoding samples during the target enrichment process enables the users to
pool multiple samples per sequencing run, and deconvolute the sample source during the data analysis step based on the barcode.
|Library Preparation Using Amplicon Tagging|
Library preparation for next-generation sequencing is by far the most time and labor consuming part of the entire next-generation sequencing process. While necessary for whole genome sequencing studies, the process can be almost entirely eliminated for resequencing projects by using PCR and amplicon tagging. By incorporating the adaptor sequences into the primer design the amplicon product is ready to go directly into clonal amplification since it already contains the necessary capture sequences.