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查看更多产品信息 TaqMan™ SARS-CoV-2, FluA, FluB RT-PCR Assay Kit - FAQs (A47701)
14 个常见问题解答
During assay validation, one should ensure that the chosen analysis parameters work for all areas of the plate. Signal can vary from one part of the plate to another, for example the corners vs. the middle.
During assay validation, one should ensure that the chosen analysis parameters, such as thresholds and Ct cutoffs, are appropriate on all instruments of the same model being used. Small variations in instrument hardware can cause differences in background signal and analysis parameters must account for all those differences
Different real-time PCR instrument models may require different threshold levels for the SARS-CoV2/FluA/FluB multiplex targets.This is due to the fact that the different instrument models have different optical designs. Therefore, during assay validation, one should ensure that chosen analysis parameters, such as thresholds and Ct cutoffs, are appropriate on all instruments to be used with the assay.
The chosen thresholds for each target must be based on comprehensive testing in your laboratory, across all instruments that will run the multiplex assay. The thresholds should be set such that they exclude the assay background fluorescence for all instruments.
Background signal can vary depending on how many positive targets are in the sample, and the sample type.
Specific analysis parameters, such as target-specific thresholds, must be determined by the user through comprehensive studies before implementation.
Using 46 PCR cycles better enables data visualization for all 3 phases of PCR (geometric, linear, and plateau) and increases endpoint fluorescence levels. This allows users to designate higher thresholds that will avoid background signal.
This incubation step enhances the mixing of reagents to avoid baseline anomalies during thermal cycling.
The volumes chosen for the extraction workflow must be validated in your laboratory before implementation.
The volumes chosen for the extraction workflow must be validated in your laboratory before implementation, even if you use the volumes recommended in the Quick Reference Card.
Yes, the workflow must be validated in your laboratory before implementation.
Any commercially available control with published concentration would be suitable for this purpose (e.g., from BEI, ATCC, etc.).
The TaqMan SARS-CoV-2, Flu A, Flu B RNA Control concentration is proprietary. This control is intended to be used as a process quality control with the TaqMan SARS-CoV-2, Flu A, Flu B Multiplex Assay. It is designed to confirm that viral targets can be routinely detected at low target levels and should not be used for workflow validation purposes.
Your chosen workflow, including extraction methodology, must be validated through comprehensive experimentation before implementation.