Precautions of PCR Tubes In Experiments
What is a PCR tube: with the highest grade of medical-grade polypropylene, PCR tubes achieve better thermal conductivity, fast response, and uniform thermal conductivity.
The key factors affecting the PCR reaction of PCR tubes:
- PCR instrument, enzymes, magnesium ions, buffers, primer annealing temperature, etc,
- Among these, the quality of PCR consumables and the temperature accuracy of PCR instruments are so close to the experimental results
- PCR tubes and PCR plates require extremely thin walls and height uniformity.
- The thickness of the PCR tube wall, and all volumes of the PCR tube, will affect the heat transfer and heat transfer rate.
- And affect the results of PCR, especially PCR (quantitative PCR) also including homogeneity and light transmittance of the tube.
- With good heat transfer, optical permeability, tolerance, and airtightness are often a priority in the selection of PCR tubes.
Precautions for experimental operation:
PCR tubes use precautions:
- High-precision small-throughput molds produce PCR consumables to ensure the uniformity and synchronization of plastic raw materials in the mold.
- Cross-contamination between samples is also one of the reasons- causes for false-positive reactions. And pay attention to all aspects of magnification.
- Wear disposable gloves. If the reaction solution is accidentally spilled, change gloves immediately.
- Use disposable tips. It is strictly forbidden to mix the PCR product analysis room with the pipette tip.
- Not exposing the tip to the air for a long time to avoid aerosol pollution.
- Avoid splashing the reaction liquid. To avoid this when opening the reaction tube, centrifuge the solution to collect the liquid at the bottom of the tube before opening the lid.
- If it accidentally spills on gloves or on the table. , immediately change gloves and wipe the tabletop with dilute acid.
When importing PCR tubes to handle multiple samples:
- Pcr tube used in multiple samples, prepare the reaction solution, first mix dNTPs, buffer, primers, and enzymes
- And then divide the package, which can reduce the operation, avoid pollution, and improve the accuracy of the reaction.
- Finally, add the reaction template, and cap the reaction tube tightly after adding.
- Set yin and yang control and blank control during operation, which can verify the reliability of PCR reaction and assist in judging the reliability of the amplification system.
- Whenever possible, use replaceable or autoclaved samplers. Since contaminated samplers with product aerosols or sample DNA, use replaceable or autoclaved samplers.
- Samplers, at least during PCR operations, should dedicate and not interchangeably, especially not carried the samplers used for PCR product analysis carried over to the other two areas.
- Repeat the experiment, verify the results, and draw conclusions with caution.