(Size: 100 rxns)
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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2/COVID-19) RT-qPCR Kit for efficient cDNA synthesis and Real-Time PCR in a single tube. This kit contains a set of primers of fluorescent probes to differentiate between 2019-nCoV/COVID-19/SARS-CoV-2 and SARS-CoV (bat SARS related CoV). SARS-CoV-2 RNA can be detected in the FAM channel and SARS-CoV RNA can be detected in the HEX, VIC or JOE channels (depending on the equipment used). Kit Contents:
- 2X qPCR Master Mix: 1 vial
- RT Mix: 1 vial
- PCR Primer/Probe Set: 1 vial
- Rehydration Buffer: 1 vial
- SARS-CoV-2/COVID-19 Positive Control (PTC): 1 vial
- Non-Template Control: 1 vial
Research Articles on Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
||Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
||Store all components at -20 °C. Avoid repeated freeze/thaw cycles.
|Directions for use
- Thaw all kit components on ice. In dark conditions, rehydrate the PCR Primer/Probe Set with Rehydration Buffer and aliquot into different tubes. Store any unused aliquots at -20 °C.
- Create a reaction mixture using the following reagents. It is recommended to multiply each of these volumes by the number of reactions being carried out, plus two additional doses. Mix the reagents well.
(For example, when carrying out 25 reactions, this would use 270 µl of Master Mix, 27 µl of RT mix, and 108 µl of PCR Primer/Probe Mix.)
|2X qPCR Master Mix||10|
|PCR Primer/Probe Mix||4|
- Set PTC, test sample and NTC wells on the microplate and record their positions. Add 15 µl of reaction mixture to each well.
- Add 5 µl of RNA extracted from samples to the sample wells. Add 5 µl of PTC to the positive control wells. Add 5 µl of NTC to the non-template control wells.
Note: The quality of the results depends on the quality of the RNA sample tested. Unsuitable collection and/or storage may give false negative results.
- Program the appropriate PCR cycling protocol on your RT-PCR instrument. A suggested thermal cycle is below.
Note: Acquisition must be performed at the end of the Annealing and Extension stages.
|Step||Temperature (°C)||Time (seconds)||Cycles |
|Annealing and Extension||58||30|
- Select the fluorescent channel (FAM/HEX) of the instrument for testing.
- 2019-nCoV/COVID-19/SARS-CoV-2: FAM (465-510 nm)
- SARS-CoV: VIC/HEX/JOE (533-580 nm)
- Follow the instrument software instructions to generate cycle threshold (Ct) values from the acquired data. Once the Ct values have been calculated from the qPCR data, the samples can be judged as follows:
- If both the FAM and HEX Ct values for a sample is at or below 35, that sample is positive.
- If both the FAM and HEX Ct values do not appear, that sample is negative.
- If the HEX Ct is at or below 35, but the FAM Ct does not appear, the sample is positive for a coronavirus, but not for COVID-19.
- If the FAM Ct is at or below 35, but the HEX Ct does not appear, the test has failed and the sample should be re-tested.
- If a Ct value above 35 appears, it is recommended to re-test the sample. If the second test appears with a Ct at or below 39, the sample is positive; otherwise, it is negative.
- The PTC should show Ct values at or below 35 for both FAM and HEX; if not, the test has failed.
- The NTC should show no Ct values for both FAM and HEX; if not, the test has failed.
|FAM / COVID-19||HEX / SARS-CoV||Outcome|
|Positive||Positive||Positive for COVID-19|
|Negative||Positive||Positive for SARS-CoV, negative for COVID-19|
|Negative||Negative||Negative for COVID-19 and SARS-CoV|
||Shipped within 5-10 working days.
||This product is for research use only.