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If an assay shows clustering around the no template control (NTC), search for an outlier sample. In some cases, the software automatically scales the plot to include the outlier giving the other samples the appearance of clustering around the NTC. If the outlier is removed from the analysis, the program rescales the plot and the data can be autocalled.
Allelic discrimination outliers
Remove the outlier(s) and reanalyze. The program will adjust the scaling and distinct clusters can be seen.
7900 SDS Software
Note: The instructions below are for Sequence Detection Software v.2.3. For other versions, see the user guide.
7500 Fast System Software
Note: The instructions below are for Sequence Detection Software v.1.4. For other versions, see the user guide.
StepOne™/StepOnePlus™ System Software
You can experience problems with the software if (select one):
The autocaller algorithm is disabled
To achieve automatic allele calling of cluster plots generated by an end point read, the automatic allele calling feature must be enabled.
Solution: How to enable the autocaller
7900 SDS Software
Note: The instructions below are for Sequence Detection Software v.2.3. For other versions, see the user guide.
7500 Fast System software
Note: The instructions below are for Sequence Detection Software v1.4. For other versions, see the user guide.
StepOne™ / StepOnePlus™ System Software 2.0
You have a low number of clusters
If only two different genotypes are present on the plate, the software will not automatically call them unless Two Cluster Calling under Function is activated.
Solution:
7900 SDS Software
Note: The instructions below are for Sequence Detection Software v.2.3. For other versions, see the user guide.
7500 Fast System Software
Note: The instructions below are for Sequence Detection Software v1.4. For other versions, see the user guide.
StepOne™ / StepOnePlus™ System Software 2.0
For Research Use Only. Not for use in diagnostic procedures.