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| 48.48 Access Array | ||
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IFC Controller AX |
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Fluidigm's Access Array System delivers the throughput and ease-of-use required to get the most out of your next generation sequencer.
Fluidigm's new Access Array is a microfluidic chip that allows the user to amplify 48 specific amplicons from 48 unique samples, in effect preparing 48 libraries in just a few hours. The power of the Access Array does not stop with massive throughput capability. Due to the unique design of the Access Array, each sample can be differentially barcoded and tagged at the amplification step, allowing for multiplexing at the sequencing step and completely eliminating the need for traditional library preparation. Access Array is an open platform and as such it can be used on any PCR-based sample preparation application using the reagents and the primers of your choice.
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Reactions |
Output |
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| 48 Samples per array | 2,304 individual PCR reactions | Product pooled automatically |
| 48 Primer pairs per array | 30nL individual reaction volume | Each sample is harvested individually |
| Only 96 pipette steps per array | Fixed reaction volumes assure reproducibility | Only 48 pipette steps to harvest product |
Sample capture and target enrichment refer 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, sample capture and target enrichment technologies allow you to select the specific areas of interest from each individual and thus only sequence that specific area of interest.
One of the largest challenges facing next-generation sequencing operators today is how to utilize the massive amounts of throughput enabled by the new crop of instruments. 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 of running multiple samples per run, and thus harvesting the tremendous throughput of these instruments. Barcoding samples during the sample capture process enables the users to combine multiple samples per sequencing run, and then later during the data analysis step, determine which sample the resulting data came from.
Library preparation for next-generation sequencing is by far the most time and labor demanding part of the entire next-generation sequencing process. While necessary for whole genome sequencing studies, the process can be almost entirely eliminated for deep resequencing projects through the use of amplicon tagging. By incorporating the adaptor sequences into the primer design the final PCR product is ready to go into emPCR or onto the flowcell since it already contains the necessary capture sequences.
| Access Array Workflow | ||
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1. Dispense samples |
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2. Dispense primers |
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3. Load the array |
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4. Thermal Cycle |
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5. Harvest the PCR product |
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6. Collect samples |
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Q – What are the minimum sample concentration and volume requirements for use with the Access Array™ IFC (Integrated Fluidic Circuit)?
A – The Access Array IFC requires a minimum sample concentration of ~50ng/µl of gDNA in order to ensure uniform sample distribution between the individual PCR reactions. Only 2.5µl of each sample is required.
Q – Is the Access Array IFC compatible with long-range PCR applications?
A – The Access Array IFC currently supports standard PCR applications but a long-range PCR protocol will be released shortly.
Q – How many samples and amplicons can I multiplex using the Access Array IFC?
A – Each Access Array IFC is capable of multiplexing 48 unique samples against 48 unique amplicons for a total of 2,304 independent PCR reactions.
Q – How much target sequence can be enriched with a single Access Array IFC?
A – A single Access Array IFC is comprised of 2,304 individual PCR reactions. Each reaction is capable of amplifying 500bp of target sequence for a total of 1.1 Mb of product per Access Array IFC. Product from multiple Access Array IFCs can be combined to increase the total amount of material per sequencing run.
Q – How does the Access Array IFC work?
A – The Access Array IFC is a unique microfluidic chip that creates 48 independent PCR reactions from each sample and amplifies a unique amplicon within each.
Q – How much genomic DNA is contained in each reaction?
A – Assuming a starting sample concentration of 50/ng per µl, each Access Array IFC reaction will contain ~200 copies of human gDNA.
Q – Can I run samples other than human?
A – Yes, the Access Array IFC is an open platform,. You are free to run any sample and primer sets you choose.
Q – What is the volume of each Access Array IFC reaction?
A – Each Access Array IFC is exactly 30nl. The accuracy of our manufacturing processes ensure that all reactions are exactly the same.
Q – What is the size range of amplicons generated with the Access Array IFC?
A – The Access Array IFC is capable of generating the exact same product size as any traditional PCR reaction. C urrently most projects use amplicons up to 500bp in length.
Q – Do you design custom assays?
A – No, the Access Array IFC is an open platform, meaning customers use their own predesigned primers.
Q – How many samples can I multiplex per sequencing run?
A – Each Access Array IFC allows you to amplify and barcode up to 48 samples, however the product from multiple Access Array IFCs can be pooled to increase the multiplexing per sequencing run.
Q – How does barcoding work?
A – Each sample is amplified with a unique set of PCR primers that contain an additional DNA sequence unique to the specific sample. Post sequencing data analysis uses the additional sequence to determine which sample the data belongs to.
Q – How can library preparation be included in target enrichment?
A – In addition to the barcoding sequence, each primer pair can also include the adaptor sequences specific to the sequencing instrument being used. The final product from the Access Array IFC includes both A and B adaptor sequences spanning the segment of interest, therefore negating the need for standard library preparation.
Q – How is the Access Array IFC different from hybridization-based sample capture technologies?
A – The Access Array IFC can prepare up to 48 samples per run. The samples can be uniquely barcoded and prepared for sequencing, therefore eliminating the need for library preparation. Hybridization-based techniques can only prepare one sample per run and do not allow for barcoding or sample preparation. This means that traditional sample preparation techniques still need to be used after sample capture.
Q – How is the Access Array IFC different from other PCR-based products?
A – The Access Array IFC is an open platform, which means the customer can use primers of their choice. In addition, other companies do not allow the samples to be uniquely barcoded or tagged with adaptor sequences. This limits the number of samples per sequencing to one, and requires a library preparation step.