Introduction

Once complete you can analyse your experiment by adding an analysis module. This section of the quick start guide will take you through the basics of Quantification and Melting analysis as well as looking at Automatic Dye File Generation.

Quantification

Both Absolute and Relative Quantification can be formed on the MyGo Mini by the using the Auto. Quant analysis module. To add this module select the “+” button in the analysis window and you will be presented with the following window:

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Analysis types are separated into Automatic and Manual sections. Automatic analysis types optimise settings suited to your data to present the best quality data. Manual analysis types give the user the option to change settings if they so wish. Automatic analysis types show what can be applied to your experiment.

Automatic Quantification

Below a 2 fold Dilution Series has been opened and an Auto. Quant. analysis module added.

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Settings

Auto. Quant. gives the user the option to exclude early cycles and alter the amplification quality. To change these values, enter your chosen setting into the fields below:

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Quantifiers

If you have assigned standards to your experiment you will be able to see how efficient your reaction is by selecting Quantifiers. Here you will be shown a standards curve for your dilution along with various information about the standards curve.

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Amplification

An amplification graph will be generated automatically in the Amplification tab on the right hand side of the screen. If you are doing more than one colour in your experiment they can be selected as shown below:

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Results as Table

A results table will also be generated automatically and an example is shown below:

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From left to right the columns in this table contain: well position, notes, well exclusion, sample, target, target type, Cq, positive and quantity. The results from this table can be exported and analyses elsewhere by selecting Export.

Melting

Melting analysis can be performed on the MyGo Mini with various modules including Automatic Tm Calling and Automatic HRM. These automatic modules will automatically generate melt peaks and cluster genotypes based on HRM analysis.

Automatic Tm Calling

Below an Auto. Tm module has been added to the experiment and the software has automatically generated the melt peaks.

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Auto Settings

The user has the option to change the target they are looking at. This would be useful if you had one set of melt peaks in some wells and another set of melt peaks in another that are using the same dye.

Signal-to-noise Ratio

The user has the option to alter the Signal-to-noise Ratio. A higher number will make the data smoother, with the risk that peaks may be missed. A smaller number will increase the opportunity of detecting smaller melt peaks.

Peaks

This window will show you the melt peaks that the module has generated. Below is an example of some automatically generated melt peaks:

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Melt peak indicators can be turned off by unchecking the Peaks selection box.

Melt

The melt can also be viewed by selecting the Melt tab as shown below:

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Automatic HRM

Automatic HRM can be performed by opening an Automatic HRM module as shown below.

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Difference Normalized and Melt

Once opened the software will automatically determine optimised parameters and generate a Difference, Normalized and Melt graph.

Clustering

This information will then be converted into a clustering graph when genotypes will be clustered based on the difference graph.

Genotypes

The software will automatically determine Inferred genotypes based on the clustering graph. However, the user can apply their own Genotypes using the Genotypes pane.

Automatic Dye File Generation

The MyGo Mini supports Automatic Dye File Generation. To create a dye file open a FSO Dye Cal. module and select the sample you wish to create a dye file from the Use Wells drop down box as shown below:

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Dye Data

This window will give you the option to choose optimised cycles for dye file generation (recommended). You can manually choose the wells you wish to create a dye file from or an entire sample. Please make sure a sample has been created, with a target assigned in Samples.

Export Calibration to File

To save your dye file, select this option. The name of the dye file as it appears in the software will be the same as it is in the Samples. The name of the file will be the name you save the file as.