The miniaturisation of molecular biology reactions can offer over 50-fold reductions in reagent usage, without impacting on accuracy – that’s a significant cost saving, regardless of the application.
What’s the secret? The key is to take advantage of the precision and reproducibility offered by automated liquid handling systems.
We’ve found that we can successfully reduce reaction volume without affecting the quality of results. This is true even for highly sensitive applications like qPCR, where minute pipetting errors are magnified at low reaction volumes.
Reaction miniaturisation for qPCR
The current challenges of using manual dispensing for qPCR can include:
- reproducibility: errors in dispensing are common when pipetting tiny volumes and have been addressed with automated dispensing systems. Errors may create false data and risks wasted time and materials in attempts to reproduce results
- accuracy: quality of data can be compromised by using manual pipettes for sub-microliter volumes, or when using automated high volume systems that are unsuitable for low volumes
- cost: buying specialised precision pipettes for low volumes, the need for frequent calibrations to ensure accuracy and the purchase of associated precision tips often proves much more expensive than using an automated system
- dead volume: large amounts of precious sample are lost due to high dead volumes
- time: it takes approximately 30 min per plate for manual pipetting but less than 10 min per plate using automated set-ups
In one recent study, we compared the qPCR results generated by dispensing reagents either by hand or using our automated liquid handler mosquito® HV. Reactions prepared using mosquito yielded comparable data sets to those dispensed by hand pipetting. However, the automated approach was faster, achieving more reproducible data while using lower sample and reagent volumes.
The automated dispensing machinery also increased statistical confidence in the results by lowering the variability between replicates. As such, this approach makes it easier to tease apart any differences between samples (Figure 1).
Figure 1: Raw data from qPCR experiment showing increased reproducibility with automated dispensing (B) compared to manual dispensing (A)
Making the most of your funding and samples
In order to keep your research programs running effectively in times of limited funding, you need to always be on the lookout for more cost and time effective ways to perform your experiments. While such optimisation has been commonplace in high throughput drug discovery labs for many years, molecular biologists are now also starting to take advantage of the benefits offered by automated systems.
This trend has been driven in large part by the rapid uptake of ‘omics’ technologies within molecular biology labs, which has greatly increased the speed and scope of data generation and analysis. This in turn has encouraged researchers to explore ways to boost throughput and reduce reagent expense.
Assay miniaturisation is an excellent solution as it enables:
- more experiments to be run simultaneously using high density multi-well plates
- expensive reagents to be used more efficiently
- precious and/or hard to obtain samples to be used more effectively
Automation makes it possible
Reducing reaction volumes is an excellent way to reduce cost, but most kit suppliers provide recommended reaction volumes for a reason; hand pipetting at low volumes across a large number of reactions is often not precise or accurate enough to generate reproducible results, not to mention being time consuming, fiddly and tedious!
Molecular biologists also routinely work with solutions off varying viscosities, from free flowing liquids such as water through to thick and sticky glycerol stocks. This further complicates the issue of accurate dispensing, especially at low volumes.
Fortunately, some modern automated systems are designed to provide exceptional precision and accuracy, even when working with liquids exhibiting a wide range of viscosities. They can be used to process many thousands of reactions quickly and effectively, freeing up your time for more important things, like analysing your results! TTP Labtech’s range of liquid handlers (including mosquito and dragonfly) use positive displacement with stainless steel pistons to offer true positive displacement pipetting. This means pipette tips never block or clog and solutions are aspirated or dispensed accurately with no dead volumes, no matter what the liquid type, viscosity, surface tension or environmental conditions.
Best of all, by providing significant cost savings, the equipment required quickly pays for itself. This allows you to produce more data much faster, all with greater accuracy and at lower cost. The system can also be applied to a wide range of molecular biology reactions such as cloning and sample prep for next generation sequencing, maximising usefulness for the whole lab.
Want to know more about how reaction miniaturisation could help make your funding go further? If so, check out our recent article on the topic over at the sp2 magazine website (pages 34-35), or take a look at the liquid handling solutions available here on our site.