Simplify your PCR pipetting strategy – tips and tricks from the industry

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Simplify your PCR pipetting strategy – tips and tricks from the industry

PCR has become a household term during the COVID-19 pandemic, though its invention and applications span back decades, where its ability to rapidly amplify target DNA sequences has been used as a fundamental technique in research, diagnostics and forensics. PCR is comprised of numerous stages, with multiple pipetting steps that often prove time consuming and troublesome. We will demonstrate how the right tools can eliminate your pipetting predicaments, vastly improving the reproducibility and productivity of your PCR workflows.    

Challenges of pipetting in PCR applications

PCR is a robust method, but it involves many pipetting steps for both reagents and samples, increasing the possibility of human errors and potentially leading to incorrect results or even failed experiments. Some common PCR pipetting challenges include:

  • Precise and repetitive procedures require high levels of concentration for long periods, where interruptions may lead to errors       
  • Temperature-sensitive reagents and samples can affect accuracy
  • Pipetting small volumes that may vary in viscosity, as well as ‘sticky’ DNA, can be difficult 
  • A variety of different labware is often compulsory, requiring the use of single-channel pipettes, leading to fatigue and increased chance of errors

Quality pipetting solutions that maximize PCR efficiency

Correct pipetting techniques and experimental set-up are well understood – and generally well practiced – across various molecular biology disciplines, and both are crucial to reducing the chances of contamination and ensuring accurate results. However, that doesn’t make it any easier to overcome the challenges discussed above – especially when large numbers of samples are involved – which require a more comprehensive solution.

At INTEGRA, we have specific pipetting technologies to address the many challenges in your PCR workflow, whatever your throughput needs. Pipette volumes range between 0.2 µl and 5000 µl, and our innovative reservoir solution – SureFlo – significantly reduces dead volumes, saving on expensive reagents. Furthermore, all our tools can easily be cleaned and sterilized, and readily fit inside biosafety cabinets to reduce the chance of contamination.

SureFlo™ anti-sealing array

Easing strain with convenient functions  

A good starting point for lower throughput PCR applications – up to half a plate per day – is our EVOLVE single or multichannel pipettes, which feature convenient volume adjustment dials to increase the accuracy and speed of manual handling. Our range of VIAFLO electronic pipettes is also suitable for low throughput PCR set-up, and perfectly capable of a higher demand of up to eight plates per day. Electronic pipettes help to reduce the risk of errors caused by repetitive pipetting, and can accurately handle the small volumes of liquid involved in PCR set-up, combining precision results with high throughput. ‘Repeat dispense’ mode, for example, offers an efficiency-boosting option for aliquoting master mixes or pipetting reagents into an entire plate.

Read how INTEGRA’s VIAFLO electronic pipettes are helping researchers at the Harvard Medical School with their PCR liquid transfers.

INTEGRA's VIAFLO electronic pipettes are available in various volumes.

Simplifying transfer between labware

PCR set-up requires transferring liquids between different labware formats – such as PCR plates, tubes and gel pockets – which is tedious, highly error prone and, over time, can cause repetitive strain injuries. Our VOYAGER adjustable tip spacing pipettes solve these age-old problems, increasing speed and eliminating transfer errors, while an ergonomic single handed operation leaves the other hand free to handle labware. See how the VOYAGER simplifies and optimizes PCR protocols involving the transfer of samples from 1.5 ml microcentrifuge tubes to 96 well plates, as well as sample addition to agarose gels for electrophoresis.

INTEGRA’s range of VOYAGER adjustable tip spacing electronic pipettes.

Higher throughput PCR applications

For productive high throughput PCR set-up – more than eight plates per day – there is a suite of options to cater for different lab sizes and budgets. The VIAFLO 96 and VIAFLO 384 channel handheld electronic pipettes, as well as the MINI 96 channel portable electronic pipette, reduce handling steps while increasing productivity and reproducibility. The MINI 96 is especially suited to PCR set-up, and can seamlessly add template DNA from a 96 well plate to a 96 well PCR plate in one pipetting step.

Discover how the INTEGRA VIAFLO 96 electronic pipette has helped Octapharma Biopharmaceuticals GmbH in Frankfurt, Germany, to cut the time it takes to perform PCR assays by more than 60 %.

MINI 96 channel electronic pipette in four different volume ranges

Automate your workflow to skyrocket throughput

INTEGRA also offers pipetting robots that can automate workflows, saving time and increasing productivity for high throughput laboratories – or even for labs that want to reduce the risk of contamination due to manual interference.

The COVID-19 pandemic has highlighted the importance of automated solutions, with laboratories placed under heightened pressure for PCR testing. Discover how the team at the Microbiology and Molecular Pathology Department at Sullivan Nicolaides Pathology in Brisbane, Australia, overcame the challenges of SARS-CoV-2 testing using INTEGRA’s ASSIST PLUS pipetting robot. This system is ideal for developing an automated PCR protocol that takes care of the multiple pipetting steps involved in the PCR workflow, helping to meet the unprecedented demand for testing. Read how this system can be used to process more samples in less time, reducing the time required to set up a 384 well qPCR plate from 1.5 hours to 12 minutes.  

Example of the ASSIST PLUS pipetting robot transferring a Mastermix into a 384 well PCR plate.

Reduce contamination

A pipette is only as good as its tips, which is why INTEGRA has developed GripTips to complement its range of PCR pipetting solutions. GripTips are produced in a cleanroom setting to prevent contamination from RNase, DNase and PCR inhibitors, and come with a purity statement to confirm conformity. They perfectly fit all INTEGRA pipetting solutions and effortlessly snap on with a unique design to ensure they never fall off or leak, reducing the risk of contamination.

 

Further reading: The top variables that can make or break your PCR success