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Applied Biosystems™ High Resolution Melt Software v3.0.1
Description
HRM analysis is a post-PCR analysis method used to identify variation in nucleic acid sequences. The method is based on detecting small differences in PCR melting (dissociation) curves. It is enabled by high-brightness, dsDNA-binding dyes used in conjunction with real-time PCR instrumentation that has precise temperature ramp control, advanced data capture capabilities, and access to software designed specifically for HRM analysis. High Resolution Melt Software v3.0.1 brings HRM to the StepOne™ and StepOnePlus™ Real-Time PCR Systems, and the 7500 Fast Real-Time PCR System, and contains features that make fast and accurate HRM results more accessible than ever.
Key product features:
• Minimal user input to achieve results—software automatically applies default/saved analysis settings upon opening an experiment file
• Easy recall—new Analysis Settings Library saves settings for future experiments
• Greater accuracy—clustering algorithm accurately distinguishes control and variant genotypes
• Increased sensitivity—software allows input of expected number of clusters for difficult-to-detect SNPs (class 4)
• Ability to achieve more in one run—separate analysis of multiple assays run on a single plate
• Shortcuts in experiment setup—direct copy and paste functions with Excel; well information and plate layout clearly displayed in Plate View
Minimize User Input to Achieve Results and Easily Recall Analysis Settings
High Resolution Melt Software v3.0.1 automatically applies default or saved analysis settings upon opening an experiment file, which allows the user to achieve results with minimal input. It also includes a new Analysis Settings Library, allowing the user to save optimized settings and easily recall them for use in future experiments.
Realize Greater Accuracy and Increase Your Sensitivity
The v3.0.1 software utilizes an improved clustering algorithm that helps to accurately distinguish control and variant genotypes, and allows the user to input the expected number of clusters for increased sensitivity for difficult SNP genotyping.
Achieve More in One Run
With the v3.0.1 software, you now have the ability to conduct separate analyses of multiple assays run on a single plate.
Get Your Experiments Up and Running Fast with Shortcuts in Setup
Users of High Resolution Melt Software v3.0.1 can copy and paste directly from Excel spreadsheets to set up experiments quickly. Plate View also clearly displays well information and the plate layout to ease plate setup.
Specifications
Specifications
| Content And Storage | Software CD, License Key |
| For Use With (Equipment) | 7500 Fast System, StepOne™, StepOnePlus™ |
| Software Type | HRM Analysis Software |
| Product Type | Software |
| For Use With (Application) | High Resolution Melting Analysis |
| Operating System | Windows 7, Windows XP Pro SP2 |
| Quantity | 10 Licenses |
Frequently Asked Questions (FAQs)
We recommend to use 50-250 bp long PCR amplicons. Typically, shorter amplicons can distinguish the genotypes for a SNP better, especially for Type III and Type IV SNPs. This is simply because a single base variation affects the melting behavior of a 100 bp amplicon stronger than of a 500 bp amplicon, for example. In longer fragments, the risk of covering multiple mutations is also increased.
Positive control reactions should contain template DNA with a known sequence. In SNP genotyping experiments, this could be a sample with a known genotype. Positive control(s) for all genotypes should be included where possible to serve as a reference in melting curve comparison and assigning genotypes for test samples. In mutation scanning experiments, a sample with a wild type sequence could serve as a positive control. The controls should preferably have the same DNA concentration as their corresponding test samples. Control DNA should also be eluted and/or diluted in the same buffer as the samples.
A 3-step cycling protocol is recommended for the analysis of complicated (especially Type IV SNP) targets, amplicons longer than 200 bp, and amplicons with a primer annealing temperature that is less than 60 degrees C.
HRM can be performed on the 7500 Fast, 7900HT Fast, StepOne/StepOnePlus instruments and requires a separate purchase of HRM software. The following instruments are also compatible with HRM and have a built-in analysis module within the instrument software: ViiA7, QuantStudio 6 and 7 Pro, QuantStudio 6 and 7 Flex, QuantStudio 12K Flex, QuantStudio 1, QuantStudio 3, and QuantStudio 5. To activate the HRM module, you will need to purchase a license.
To perform an HRM experiment, you will need the reagents to calibrate your instrument for the HRM dye and the software to perform the analysis.