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  9. Advantages of Applied Biosystems HRM

Advantages of Applied Biosystems HRM

Identify More New Variants, Quickly and Accurately.


Difference plot generated on another plate-based real-time HRM system

Figure 1. Difference plot generated on another plate-based real-time HRM system.

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 ViiA™ 7, 7900HT Fast, 7500 Fast, StepOnePlus™, and StepOne™ Real-Time PCR Systems. The StepOne™ (48-well) and StepOnePlus™ (96-well) Real-Time PCR Systems are remarkably simple systems designed with a user-friendly, yet powerful, interface for researches of all experience levels. 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 ViiA™ 7 and 7900HT, the gold standard high-throughput systems. 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.

The ability to easily identify new variants is key for successful HRM applications. An HRM system from Competitor R (Fig 1) was only able to distinguish them in 10% of the population.

By eliminating the temperature shift step, the Applied Biosystems HRM solution (Fig 2) was able to clearly distinguish homozygous variant samples from homozygous wild type samples in 97.5% 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).

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 with the Applied Biosystems HRM application on the 7500 Fast and 7900HT Fast Real-Time PCR Systems.

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