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PCR Setup: What You Need to Know

Image of electrophoresis gel and PCR curves for article on PCR.

PCR tests have long been used to see genetic changes and diagnose infectious diseases. The test takes many copies of a DNA sample, allowing researchers to obtain a small sequence of DNA that can be amplified to study the DNA in greater detail. While this type of test became widely known during the COVID-19 pandemic and beyond, PCR tests have a variety of uses.When it comes to manual vs automated PCR handling, there are quite a few benefits to using a semi- or fully-automated PCR robot. Automation not only speeds up test results but has other benefits as well. Read on to learn more about PCR setup and why robotic PCR handlers are a much better option for extraction.

Manual vs Automated PCR Testing

Even though automation is a more reliable way to isolate and extract RNA and DNA, some clinical laboratories still use manual testing. Traditional PCR testing isn’t an incorrect method but can be more fallible than automated PCR testing for several reasons and poses some challenges.  First, manual testing requires human manipulation of the sample and reagents, which increases the risk of contaminating the sample and exposing doctors and researchers to potential infection, depending upon what’s being tested.

The consistency of the isolated nucleic acid is maintained using a robotic handler, as fewer manipulations are needed for the sample. While this prevents cross-contamination, it also promotes efficiency, as much less manual labor is involved. In addition, more laboratory professionals can be present for testing because of automated extraction instruments.

Smaller labs may opt for manual PCR testing because of the cost of automation. However, a fair amount of throughput of samples justifies the cost and expedites the process. And, samples have much less risk of contamination and are more accurate.

Setting up for Automated PCR Testing

Setting up for automated qPCR and PCR robots is quite simple. There are machines designed for liquid handling and PCR automation that are relatively customizable and composable, targeting a laboratory’s needs. Simple PCR robots have simple, easy-to-read touchscreen controls and can be set up in minutes. However, any other throughput needs can be addressed, and robot handlers can be designed for PCR setup with an integrated thermocycler, optimal normalization, hands-free sample transfer, and DNA and RNA isolation, which is all optimal for either endpoint, real-time, RT, or multiplex PCR experiments. Platform sizes can range from benchtop (simple PCR robots) to integrated, large work areas.

PCR robots combine samples and PCR mastermix in the reaction plate, which goes into a qPCR machine or thermocycler. The mixture is prepared beforehand by either using the robot or by hand (more of a semi-automated process). This PCR robot works well for gene expression (qPCR), NGenotyping, cloning amplification, and medical, applied, and forensic sciences. Automated PCR testing works by:

  • Using the software aligned with your PCR robot, use a template from a protocol library, or design your own.
  • Have the machine make the mastermix or make it by hand.
  • Combine mastermix with forward and reverse primers.
  • Calibrate and activate the protocol with touchscreen controls.
  • Load the thermocycler and run the machine.

Having the machine prepare the mastermix for you further reduces the risk of contamination or cross-contamination.

Benefits of Automated PCR

In addition to streamlining the process and reducing the risk of contamination, there are other benefits to using PCR robots. Some of the advantages include:

  • High throughput. The more hands-off the PCR process is, the higher throughput you can attain. Microplate and liquid handling robots help accomplish this.
  • Reduces liquid waste. Using an automated reagent dispenser, you can control the amount of liquid that enters each pipette. This reduces reagent waste and saves time.
  • Sample purity. Integrating a magnetic bead extraction machine increases throughput and the purity of the sample to avoid false positives or negatives.
  • Improves traceability. Samples and processes in the laboratory should be completely traceable, and lab automation software with a PCR robot helps accomplish this.
  • Improves reproducibility. Standardized PCR controls help improve reproducibility within the process.

Opting to take the laboratory semi- or fully automated with a PCR robot has many advantages and very few drawbacks. Its utilization helps you keep up with modern-day standards while reducing contamination risk and providing more pure samples with higher throughput.

Hudson Robotics has been the leading supplier of automated products for over 38 years, including many aspects of PCR automation. Speak to a representative today!