The HibriGen COVID-19 RT-PCR Kit is used for the qualitative detection of the SARS-CoV-2 (2019-nCoV) virus by RNA obtained from human nasopharyngeal aspirate / lavage, bronchoalveolar lavage, nasopharyngeal swab, oropharyngeal swab, and sputum samples.
The method of operation of the kit is the single-step RT-PCR (real-time fluorescent PCR) technology, which collects the cDNA mapping (Reverse Transcription) from the RNA of the SARS-CoV-2 (2019-nCoV) virus and the real-time amplification (RT-PCR) of the cDNA in one step.
Primers and probes included in the HibriGen COVID-19 RT-PCR Kit are specific to sequences of the protected RdRp gene (FAM), N gene (Hex) and Human RPII gene for reaction internal control (Cy5) as target sites. It has been designed in accordance with RT-PCR technology from a single step.
50 Test |
100 Test |
200 Test |
500 Test |
1000 Test |
2000 Test |
|
2X One Step RT-PCR Mix |
1×500 ul |
1×1 ml |
2×1 ml |
5×1 ml |
10×1 ml |
20×1 ml |
5X Primer Probe Mix |
1×200 ul |
1×400 ul |
1×800 ul |
2×1 ml |
4×1 ml |
8×1 ml |
For 20 pl total reaction volume;
Content | Amount | Final Concentration |
2X One Step RT-PCR Mix | 10 ul | 1X |
5X Primer Probe Mix | 4 ul | 1X |
Sample | 6 ul |
1-The prepared RT-PCR sample is gently mixed and centrifuged briefly to collect
the remaining drops on the walls of the tube completely to the bottom of the tube.
2-In PCR application, apply the following thermal cycle conditions.
Number of Cycle |
Temperature |
Duration |
1 |
55°C |
15 Min |
1 |
95°C |
1 Min |
45 |
94°C |
5 Sec |
60°C |
20 Sec |
|
FAM/HEX/Cy5 reading |
FAM |
HEX |
CY5 |
Interpretation |
+ |
+ |
+ |
Positive |
– |
– |
+ |
Negative |
+ |
+ |
– |
Unefficient RNA isolation / Positive |
– |
+ |
+ |
Positive |
+ |
– |
+ |
Positive |
– |
– |
– |
Repeat |