Affymetrix Human SNP 6.0 Genome-Wide Genotyping Services

Single nucleotide polymorphisms (SNPs) are the most common type of genetic variation and represent over 80% of the genetic variation between individuals. SNPs are ideal candidates for research correlating phenotype and genotype. Since some SNPs predispose individuals to a certain disease or a trait or cause an altered reaction to a drug, they are proving to be highly useful in diagnostics and drug development.

With more than 1.8 million genetic markers, Affymetrix’ SNP 6.0 array provides high-performance, high-powered and low-cost genotyping. It is now available from Asuragen. In combination with Asuragen’s service expertise you have the tools to carry out a whole-genome study and bring power to your research. Asuragen performs hundreds of Affymetrix profiling jobs per month, and was the only commercial service provider participating for the Affymetrix platform in the pivotal MAQC study. Our validated procedures and optimized processes give you the quality and turn-around time critical for a successful experiment. Learn more about how we qualify our methods and staff.

Description of Services

Asuragen provides services using the Genome-Wide Human SNP Array 6.0, a single array with more than 1.8 million markers of genetic variation, including more than 906,600 SNPs and more than 946,000 non-polymorphic probes for the detection of copy number variation. The SNP Array 6.0 demonstrates excellent performance and represents more genetic variation on a single array than any other product, providing maximum panel power and the highest physical coverage of the genome (median marker spacing of 680 bases).

The SNPs were chosen from the public and Perlegen SNP databases and are represented on this array based upon reproducibility, call rate, accuracy and concordance with genotypes from the International HapMap Project (for more information on SNP selection – link to Affymetrix website). Of the 946,000 non-polymorphic copy number probes contained on the Array, 202,000 probes were selected based on known copy number changes reported in the Toronto Database of Genomic Variants (DGV). The rest of the probes were chosen to be evenly spaced along the genome to enable the detection of novel copy number variation.

Workflow for SNP 6.0 genotyping services:

  • Incoming DNA QC (click here for sample prep services):
  • Target preparation, followed by Affymetrix recommended in-process QC
  • Array hybridization, washing, and scanning followed by array QC
  • Data summarization (see Deliverables Section)

Target preparation:

For target preparation Asuragen uses the Affymetrix validated Genome-Wide Human SNP Nsp/Sty Assay. In short, 500 ng of total genomic DNA is digested with Nsp I and Sty I restriction enzymes creating cohesive 4 bp overhangs. Universal adaptors are then ligated to all fragments resulting from restriction enzyme digestion, regardless of size, via these cohesive 4 bp overhangs. A generic PCR primer that recognizes the adaptor sequence is used to amplify adaptor-ligated DNA fragments from both restriction digests. The PCR conditions have been optimized to preferentially amplify fragments in the 200 to 1,100 bp size range. PCR amplification products for each restriction enzyme digest are combined and purified using polystyrene beads. The amplified DNA is then fragmented, labeled, and hybridized to a SNP Array 6.0 (see Figure below).

Nsp/Sty Assay

Deliverables

Included in the price of SNP genotyping, you will receive:

Analyses performed are:

  • Genotype calls produced using the Birdseed v2 algorithm, developed by the Broad Institute
  • Copy number variation analysis based on the Canary algorithm listing NC state calls. Annotations listed from the DGV

Affymetrix SNP 6.0 Deliverables:

  • Affymetrix Raw Data Files
    • Flat Files
    • CEL Files
  • Analysis Report (in MS Excel Format, unless table dimensions exceed MS Excel capabilities, in which case .txt files will be provided)
    • Project Description
    • Intensity QC
    • Genotype Summarys
    • Filtered SNP List
    • SNP Summary Statistics
    • Copy Number QC
    • Copy Number Segment Summary
    • CHP, CNCHP, CNVCHP files
  • Figures:
    • Karyoview of each sample representing cytobands depicting gains and losses for each chromosome (.pdf format)

To ensure security, all information is provided in an electronic form on a DVD. Encryption is available on request. Data may also be delivered via an FTP site or e-mail.

Quality Control

During our processes, we monitor many QC checkpoints as recommended by the Affymetrix procedures. We utilize in-process sample controls, and process or reagent batch controls. Should any sample-specific QC measure result in an “alert” or “fail” reading, we will contact you to discuss available options.

Asuragen’s rigorous procedures enable Asuragen to be the only service provider to offer a Data Quality Guarantee:

If, for any reason, a qualified sample does not pass our process control specifications, we will re-run the sample at no additional charge.

Read more about our commitment to the best possible data quality, or read about our contributions to in-process controls and standardization.

Sample Handling and Processing

All samples are received and entered into the analysis queue under a controlled SOP to confirm package/sample integrity, appropriate shipping conditions (frozen samples/dry ice, as appropriate), and individual confirmation of sample identity against the shipping manifest and sample submission form and instructions.

Turn-around Time

Most projects are completed in two weeks or less. If you have a critical time requirement, we can often meet your need for no additional charge. Read more about our commitment to prompt turn-around times.

Additional Services Available from Asuragen

Available Genome-wide SNP Arrays

Human SNP Array 6.0

Catalog Numbers for Affymetrix SNP 6.0 Services

CA0068 QC, Nanodrop and Agarose Gel, for DNA applications
CA0092 Target Preparation, Affymetrix SNP 6.0
CC0380 Affymetrix Genome-Wide Human SNP Array 6.0
CA0093 Array Processing and Scanning Affymetrix SNP 6.0; per sample

*Asuragen supports a broad range of arrays from Affymetrix. Contact Asuragen for more detail.

Sample Input Requirements for the services listed above are found in the Sample Submission Form and Shipping Instructions

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