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qPCR High Resolution Melt Assays

Illumina qPCR High Resolution Melt (HRM) Assays are custom-designed, bioinformatically validated, genotyping assays composed of forward and reverse PCR primers. These assays are designed to be used with a double stranded DNA-Binding dye, such as SYBR Green. Illumina qPCR HRM Assays are designed to detect sequence variants (such as SNPs), using an advanced algorithm to ensure assay success.

qPCR High Resolution Melt Assays Workflow

Illumina qPCR HRM Assays offer a simple workflow that reduces the time you spend designing and setting up HRM genotyping experiments. Assays are created using Illumina’s custom online design tool, DesignStudio. Each assay is delivered as a ready-to-use 20X assay mix, minimizing pipetting steps. Illumina qPCR HRM Assays are designed to be used with HRM optimized DNA binding-dye master mixes.

DesignStudio Workflow For qPCR High Resolution Melt Assays

You will begin the design process by creating a project, choosing a species and choosing a reference genome build. The next step is to define target regions by rsID, chromosome coordinates, or sequence. Target regions can also be defined by uploading a file in .csv format. When you input chromosome coordinates, you can choose which type of sequence variants (SNPs, INDELS or All) you would like DesignStudio to design assays against. When you input sequence, you must include one SNP indicator in the format: [C/G]. Once your assays are designed, you can visualize their composition (including primer sequences) and position by clicking on the target name. Select the assays you want and click Add to Design. Finally, review your selected assays on the Review Design tab and click Request Quote to receive pricing for your project.