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Main Components and Contents:
| Componentes | ED2304001-01（24T） | ED2304001-02（50T） |
| Fluorescent PCR solución de reacción | 500μL per tube × 1 tubo | 1200μL per tube × 1 tubo |
| Positive control | 50μL per tube × 1 tubo | 200μL per tube × 1 tubo |
| Control negativo | 50μL per tube × 1 tubo | 200μL per tube × 1 tubo |
| Manual | 1 | 1 |
Function and Purpose:
Used for detecting target nucleic acids in various samples including whole blood, suero, lymph nodes, spleen, muscles, y otros.
Dosage and Determination:
1 Instrucciones de uso
1.1 Procesamiento de muestras
Process the samples according to relevant standards and store the processed samples for later use.
1.2 Procedimiento experimental
1.2.1 Reagent Preparation Area
Remove the reagent kit, take out the required reagents for the experiment, thaw and mix thoroughly, then briefly centrifuge for 5 seconds to remove any liquid adhering to the tube walls;
Add 20μL of fluorescent PCR reaction solution into each PCR tube and transfer it to the sample preparation area.
1.2.2 Área de preparación de muestras
Add 5μL each of the negative control, the nucleic acid of the sample to be tested, and positive control. Briefly centrifuge for 5 seconds and transfer to the amplification area.
1.2.3 Amplification Area
Place each PCR tube in the corresponding positions of the instrument’s sample slots and record the placement sequence.
Set the instrument’s nucleic acid amplification parameters according to the table below and perform the PCR amplification.
| Sistema de reacción | 25μL Reaction System | ||
| Signal Channel | FAM Channel for Collecting Fluorescent Signals | ||
| PCR Reaction Conditions | Stage | Condiciones | Ciclos |
| Pre-Denaturation | 95℃：3mín. | 1 | |
| PCR | 95℃：15segundo | 40 | |
| 60℃：30segundo | |||
2 Interpretación de resultados
2.1 Establishment of Experimental Conditions:
2.1.1 Positive Control: Valor CT ≤ 30, showing clear exponential growth, displaying a typical “S” curve.
2.1.2 Control negativo: valor ct > 38 or no Ct value, showing no clear exponential growth phase or plateau.
2.2 Criteria for Determination:
2.2.1 Positivo: Sample detection result Ct value ≤ 35, displaying clear exponential growth, indicating the presence of the target DNA in the sample.
2.2.2 sospechoso: Sample detection result Ct value between 35 y 38; the sample should be retested. If the repeated experiment results in Ct values still between 35 y 38 with clear exponential growth, se considera positivo; de lo contrario, it’s negative.
2.2.3 Negativo: Sample detection result Ct value > 38 or no Ct value, indicating the absence of the target DNA in the sample.
Precauciones:
- Before the experiment, carefully read through this reagent kit manual and strictly follow the operational procedures.
- Store all reagents at the specified temperatures; detailed information is available on the reagent labels.
- Sample handling should occur in a biosafety cabinet. After the experiment, subject all samples and materials used to high-temperature sterilization.
- To prevent cross-contamination, work in an environment as free from nucleases as possible, and segment the experimental process (Área de preparación de reactivos, sample preparation area, amplification area, etc.).
- When preparing the PCR reaction system, try to avoid bubble formation. Before amplification, check if reaction tubes are tightly sealed to prevent leakage and contamination of the instrument.
- This kit can be used with standard nucleic acid extraction purification methods for sample testing, maintaining constant judgment parameters such as Ct values.
- To ensure accurate experimental results, samples should be freshly collected and transported at 2-8°C; for long-distance transport, use dry ice.
- Avoid repeated freezing and thawing of all reagents.
Especificaciones: 24 t/caja, 50 t/caja
Storage and Shelf Life: Conservar a -20°C, away from light. Shelf life is 12 meses.
