High Melting Resolution Software for Variant Screening on Real-Time PCR Systems
           
7500 Fast Real-time PCR System

Applied Biosystems 7900 HT Fast Real-Time PCR System

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No Temperature Shift Required.
Fast. Accurate.

Identify more new variants, quickly and accurately.
High Resolution Melting (HRM) analysis is an alternative to dHPLC sequencing screening of new gene variants. The HRM Software is now available on the Applied Biosystems 7500 Fast System and on the 7900HT Fast Real-Time PCR System. The 7500 Fast Real-Time PCR System delivers precise results with fast thermal cycling in a standard 96-well format. Achieve high-throughput HRM analysis with the 384-well 7900HT, the gold standard high throughput system. The AB HRM application does not require temperature shifting, which results in a greater likelihood of identifying new homozygous mutations than methods that require temperature shifting.



New to HRM? Download our Guide to High Resolution Melting (HRM) Analysis

The Applied Biosystems HRM Software provides an easy and intuitive workflow that:
  • Shortens analysis time by auto-calling genotypes and automatically omitting the no template controls
  • Minimizes subjective analysis by automatically grouping unknown variant clusters
  • Allows easy data review with customizable plot views, expandable windows, and one-click color assignment to highlight curves of interest
  • Ability to analyze multiple targets (assays) on one plate

The Applied Biosystems HRM Advantage


 A. 7500 Fast Real-Time PCR System B. 7900HT Fast Real-Time PCR System


   Heterozygote    Homozygous - Wildtype    Homozygous - Variant

Figure 1. Difference plot generated with the Applied Biosystems HRM application on the 7500 Fast and 7900HT Fast Real-Time PCR Systems.


Figure 2. Difference plot generated on another plate-based real-time HRM system.		 The ability to easily identify new variants is key for successful HRM applications. By eliminating the temperature shift step, the Applied Biosystems HRM solution (Fig 1) was able to clearly distinguish homozygous variant samples from homozygous wildtype samples in 97.5% of the population, whereas the other HRM system from Competitor R (Fig 2) was only able to distinguish them in 10% of the population. All genotypes were auto-called by the respective software packages and were not altered by the operator. Class 1 SNP (A/G), multiple technical replicates of nine DNA samples representing three genotypes: homozygous wildtype (G/G), homozygous mutant (A/A) and heterozygous (A/G).