TaqMan™ SARS-CoV-2, FluA, FluB RT-PCR Assay Kit - FAQs

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

Is there variability in signal within a plate when using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit?

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.

When using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit, can I use the same model, but different instruments (e.g., two QuantStudio 5 instruments)?

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

When using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit, can I use the same target-specific thresholds for this assay when using different real-time PCR instrument models (e.g,. QuantStudio 5 vs. 7500 Fast Dx)?

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.

When using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit, how can background be accounted for when determining thresholds?

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.

When using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit, what does background signal look like for this multiplex assay?

Background signal can vary depending on how many positive targets are in the sample, and the sample type.

What are the analysis parameters for the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit?

Specific analysis parameters, such as target-specific thresholds, must be determined by the user through comprehensive studies before implementation.

Why does the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay workflow recommend 46 PCR cycles?

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.

What is the purpose of the 85 degrees C incubation step in the thermal cycling parameters when using the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit?

This incubation step enhances the mixing of reagents to avoid baseline anomalies during thermal cycling.

Do I have to extract RNA from 400 µL of specimen, or can I use less (e.g., 200 µL) when using with the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit?

The volumes chosen for the extraction workflow must be validated in your laboratory before implementation.

If I use an RNA extraction elution volume different from what is recommended in the TaqMan SARS-CoV-2, Flu A, Flu B Multiplex Assay - Quick Reference Card (Pub. No. MAN0019600), should I adjust the volume of MS2 added to the sample processing plate?

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.

If I follow the TaqMan SARS-CoV2, Flu A, Flu B Multiplex Assay - Quick Reference Card (Pub. No. MAN0019600) as written, do I still need to validate the workflow before implementation?

Yes, the workflow must be validated in your laboratory before implementation.

What control can I use to validate the limit of detection (LoD) for the TaqMan SARS-CoV-2, Flu A, Flu B multiplex assay ?

Any commercially available control with published concentration would be suitable for this purpose (e.g., from BEI, ATCC, etc.).

What is the concentration of the TaqMan SARS-CoV-2, Flu A, Flu B RNA Control material in the TaqMan SARS-CoV-2, Flu A, Flu B RT-PCR Assay Kit?

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.

The TaqMan SARS-CoV-2, Flu A, Flu B Multiplex Assay - Quick Reference Card (Pub. No. MAN0019600) refers to RNA extraction using the KingFisher Flex 96 System. Can I use a different RNA extraction method?

Your chosen workflow, including extraction methodology, must be validated through comprehensive experimentation before implementation.