QuantStudio™ Absolute Q™ 数字 PCR 系统，台式计算机

Continuous melt curve: increases the temperature by the ramp increment (degrees C/sec) so that the temperature will change at a constant rate set in the protocol.
Step and Hold melt curve: increases the temperature by the ramp increment (degrees C) and then holds at that temperature for the time specified by the user.

The amount of data collected will depend on the filter(s) selected. The more filters you use, the longer the acquisition will take and the fewer data points will be collected.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

The QuantStudio Absolute Q Digital PCR Instrument (Cat. No. A52864) uses QuantStudio Q MAP16 Digital PCR Plates that contain 16 digital PCR microreaction chamber arrays, each containing 20,480 fixed volume microreaction chambers of 432 ρL (0.000432 µL). If there is 1 copy of gene target in the tested sample, and the sample is using 1 array of the MAP16 plate, then the theoretical lower limit of detection (lowLOD) for the Absolute Q can be approximated by the following calculation:

lowLOD for Absolute Q = (1 copy)/ (0.000432 µL X 20,480 chambers) = 0.113 copies per µL

If a sample is run in triplicate, and there are 3 positive chambers, the lowLOD in practice for Absolute Q can be calculated using the following calculation:

lowLOD for Absolute Q = (3 copies)/ (0.000432 µL X 20,480 chambers) = 0.339 copies per µL

The QuantStudio Absolute Q has the sensitivity of detecting the presence of a single copy of target sequence within the 9 µL reaction volume, which would result in a detected concentration of ~0.1 copies per µL. This detection limit can be reduced even further by using the software’s digital pooling feature to increase the total analyzed volume. The ability of the instrument to detect at this level will be impacted by various components including the assay efficiency and reagents used in the reaction. The LOD is also dependent on a given assay’s specificity as well as the potential background fluorescence signal from the sample matrix, which may affect the LOD achieved in practice. The Absolute Q software’s unique false positive detection capability can enhance an assay’s performance, especially when low target concentrations are investigated.

It is important to note that the LOD should be determined empirically by the user for their specific experimental conditions.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

QuantStudio Absolute Q Digital PCR System (Cat. No. A52864) supports the following dyes: FAM™ dye, VIC™ dye, HEX™ dye, ABY™ dye, ROX™ dye, CY™5 dye, and JUN™ dye.

Please refer to page 13 of the QuantStudio Absolute Q Digital PCR System User Guide for Installation, Use and Maintenance for more information on supported optical dyes and channel selection.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

If dye channels are turned off before the run, then data will not be collected for them as the run progresses. This means that changing the dyes after the run will not be possible.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Unused dye channels can be turned off after the run by going to the SETUP tab and clicking EDIT SETUP and then EDIT GROUPS. Here, the Analysis can be changed to “Not Used.” After clicking SAVE twice, the data will be reanalyzed and the unused dye channels will not be present in the analysis.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.