| | The Mapping 500K Array Set is comprised of two arrays, each capable of genotyping on average 250,000 SNPs. |
| | One array uses the Nsp I restriction enzyme (~262,000 SNPs), while the second uses Sty I (~238,000 SNPs). |
| | Total genomic DNA (250 ng) is digested with a restriction enzyme (Nsp I or Sty I) and ligated to adaptors that recognize the cohesive four base-pair(bp) overhangs. |
| | All fragments resulting from restriction enzyme digestion, regardless of size, are substrates for adaptor ligation. |
| | A generic primer that recognizes the adaptor sequence is used to amplify adaptor-ligated DNA fragments. |
| | PCR conditions have been optimized to preferentially amplify fragments in the 200 to 1,100 bp size range. |
| | The amplified DNA is then fragmented, labeled, and hybridize to a GeneChip Human Mapping 250K Array. |
| | Each consisting of more than one milion copies of a 25-bp oligonucleotide probe of a defined sequence. |
| | Two reagent kits were developed and validated for use in conjunction with the Mapping 500K Array Set. |
| | One kit is specific to the Nsp I restriction enzyme while the other is designed for the Sty I restriction enzyme. |
| | Both kits contain validated and qualified reagents for the most critical steps in the GeneChip Mapping Array. |
| | This includes the PCR primer and adaptor necessary to selectively amplify a portion of the human genome, reagents to fragment and label the PCR product, and several control reagents. |
| | Kits are available for either 30 or 100 reactions. |
