MaxDirect Plus RT-PCR – RNA Extraction FREE
The Difficult RNA Extraction Step Is Gone With DIRECT RT-PCR Amplification.
Save MONEY and TIME!
SARS-CoV-2 can be detected with this Direct RT-PCR MasterMix (COVID- 19).
Catalog Number: MS13
Size: 100 tests/rxns
Also Known As: COVID-19 Direct PCR, SARS-CoV-2, Direct RT-qPCR
Description:
MaxDirect Plus RT-PCR Master Mix is capable to detect SARS-CoV2 directly from samples. The RNA Extraction Kit is NOT needed anymore. MaxDirect Plus RT-PCRs works directly from RNA templates.
Storage:
| Componentes | |
| MaxDirect Plus Plus RT-PCR 2x Probe Master Mix | -25 °C and -15 °C |
| 10x S-CoV2 Duplex Assay | -25 °C and -15 °C |
| S-CoV2 positive control | -25 °C and -15 °C |
| Direct Booster solution | 4 and 12 °C |
Shipping temperature:
Although this Master Mix is stable at +20°C for up to 9 days (if not opened). We strongly recommend using 2kg. of ice packs to keep the Master Mix in optimal conditions (no heavy dry ice needed).
MaxDirect Plus RT-PCR Probe 2x Master Mix will not lose activity within 10 freeze-thaw-cycles. Additional freeze-thaw cycles could affect the Master Mix’s performance. Keep the kit out from lighting environments.
Kit Components:
| Component | Content | Volume |
| MaxDirect Plus Plus RT-PCR 2x Probe Master Mix | DNA Polimerase, dNTP, buffer, PCR additives | 1,5 ml
|
| 10x S-CoV2 Duplex Assay | Primer/Probe Mix for SARS-CoV-2 N-gen and RNaseP | 300 µl |
| S-CoV2 positive control | DNA Plasmid with target region for SARS-CoV-2 and target region for RNaseP | 70 µl |
| Direct Booster solution | Additives that improve the sensitivity of PCR Direct | 100 µl |
PCR Set Up For ALL Sample Collection Methods:
| Step | Temperature | Duration | Cycles |
| Reverse Transcriptase | 70 °C | 10 min. | 1 |
| Denaturation | 95 °C | 5 seg. | 45 |
| Annealing | 65 °C | 20 seg.* |
- Fluorescence measurement in FAM and Cy5-channel
Thaw kit components on ice, then mix and centrifuge all components.
For the preparation of the PCR Master Mix, the number of total reactions, including positive and negative controls, is calculated. It is recommended to prepare an additional reaction to compensate for the loss of pipetting. Mix and centrifuge the components.
1st. OPTION (DRY SWAB based sample collection)
MaxDirect Plus RT-PCR Amplification Protocol
Put the dry swab stick thoroughly in 350 µl RNase-free water. Remove debris from the swab by short centrifugation. Then heat the samples for 5 min at 95 °C. The assay detects SARS-CoV2 N1-gene (FAM- channel) and human RNaseP-gene (Cy5-channel).
Assay setup
| Reagente | Volumen |
| MaxDirect Plus Plus RT-PCR 2x Master Mix | 15 µl |
| 10x Duplex Assay | 3 µl |
| PCR-Clean wáter | 5.1 µl |
| Direct Booster solution | 0.9 µl |
| Dry swab RNA sample/positive control/negative control | 6 µl |
| Total | 30 µl |
The positive control can be used directly or diluted 1 to 10, which will give a solid signal.
2nd.OPTION(GARGLE based sample collection)MaxDirect Plus RT-PCR amplification protocol.
Gargle with 5 ml of Clean Water for PCR or 0.9% saline solution for at least 20 seconds and then spit into a PCR tube.
Gargle specimens can be inactivated by heating the tube for 3 min at 95 ° C, the heat inactivated gargle solution can be stored for a maximum of 3 hours at 0-12 ° C before use. But application is recommended immediately after heat inactivation. Take proper precautions against aerosol formation during the gargling process.
Note.- Make sure the patient does not eat, drink, chew gum, or smoke for at least 30 minutes before collecting the sample.
Assay setup for Gargle based collection samples:
| Content | Volumen |
| MaxDirect Plus Plus RT-PCR 2x Master Mix | 15 µl |
| 10X Duplex Assay | 3 µl |
| PCR-Clean Water (Agua de PCR) | 5.1 µl |
| Direct Booster Solution | 0.9 µl |
| Gargling solution/positive control/negative control | 6 µl |
| Total | 30 µl |
3rd.OPTION (RNA Extraction based method)
MaxDirect Plus RT-PCR Master Mix Protocol for extracted/purified RNA samples
Follow the instruction of your RNA extraction kit to process the samples.
The assay detects SARS-CoV2 N1-gene (FAM-channel) and human RNaseP-gene (Cy5-channel).
Assay setup
| Reagent | Volume |
| MaxDirect Plus Plus RT-PCR 2x Master Mix | 10 µl |
| 10X Duplex Assay | 2 µl |
| PCR-Clean water | 3 µl |
| RNA extracted sample | 5 µl |
| Total | 20 µl |
Interpretation of data
| FAM – N1 | Cy5 – RNaseP | Interpretation |
| + | – | Unclear, repeat experiment |
| + | + | Valid – SARS-CoV2 positive |
| – | + | Valid – SARS-CoV2 negative |
| – | – | Invalid, repeat experiment |
General considerations
- Use sterile, DNA/DNase/RNase-clean consumables to avoid contaminations.
- Clean all surfaces and equipment in direct environment before starting the experiment to lower the risk of nucleic acid-, DNase/RNase- and protease contaminations.
- Do not use this product after its expiry date.
- Include positive and negative controls in your experiment to validate your results.
Our RT-qPCR Kit Is Fully Compatible With:
Different kind of PCR Instruments. Please send us an email to: info@maxprecisionlab.com and we will check the compatibility of your PCR instrument with this kit.
We Have Different Kind Of ROX Availabe:
- Low ROX,
- High ROX
-
NO ROX
Materials and Devices Required but Not Provided:
- Biological Cabinet
- Desktop centrifuge (suitable for 96-well plate or 8-strip tube).
- Adjustable pipetts (with maximum capacity of 2μL, 10μL, 50μL, 100μL, and 200μL, respectively)
- Disposable pipette tips with filters
- Disposable powder-free
- Water Bath
- Positive/Negative Controls
- RNase-free water
- PCR reaction tube / PCR reaction
NOTE: Please ensure that the PCR instrument and materials have been installed, calibrated, checked, and maintained according to the manufacturer’s instructions and recommendations.
Our Production Team Has Been Working Very Hard To Fulfill The High Demand For Covid-19 RT-PCR Kits.
Therefore The Number Of Samples Is Limited.
In Order To Confirm The Number Of Samples Available. Please Send Us An E-Mail To: info@maxprecisionlab.com
MaxSARS2 RT-PCR MasterMix
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Shipping temperature: This kit can be shipped with Ice Pack
MaxPrecision Lab Warranties that all products will be FREE from defects in materials, workmanship and will perform in conformance with our protocols*.
