miniaturisation of genomics reaction volumes
Many of the fundamental molecular biology tools and gene cloning techniques are used routinely in a wide variety of academic research and drug discovery applications.
Almost all genomics protocols, including library preparation for next generation sequencing (NGS) or simple PCR and qPCR reactions, recommend volumes that are within the dispensing range of manual pipettes, or that of larger volume liquid handlers. This ensures the optimum accuracy and precision required for these reactions.
genomics application areas for miniaturised liquid handling
TTP Labtech’s automated liquid handlers, the mosquito HTS, mosquito X1 and mosquito HV, have revolutionised the sample prep workflows in many areas, including cancer and stem cell research, through high-throughput single cell analysis, using DNA or RNA sequencing.
Validated applications from major genomics collaborators include:
- library preparation for single cell DNA and RNA sequencing
- reverse transcription using the Ultra SMARTer kit
- automated precise and accurate low-volume DNA normalisation
- PCR and qPCR reaction set-ups
- gateway cloning
- colony spotting
TTP Labtech’s liquid handlers are not restricted to specific kits or processes. mosquito liquid handlers, work similar to multi-channel pipettes but in an automated and low volume range (nL-µL) at high precision and accuracy. Being an open platform, these systems are commonly used in any enzymatic reactions that would benefit from lower reagent costs, standardized and reproducible high throughput set-up and/or reduced sample requirements. Moreover, our experts are available to help with different genomics applications, including but not limited to: NGS library prep, SPRI- bead clean-up, CRISPR-cas9 and synthetic biology applications.
which liquid handler best fits my applications?
Using low-volume liquid handling in normalisation and sample prep of genomics applications is especially beneficial when working with precious and low abundance samples such as DNA/RNA derived from patient specimens or single-cell analysis.
mosquito HTS (25 nL – 1.2 µL) and mosquito HV (0.5 – 5 µL) are automated 8- or 16-channel liquid handlers. Being based on true-positive displacement pipetting technology they enable fast, accurate, gentle and contamination-free liquid transfer, essential for genetic analysis applications where viscose gDNA and reagents are being handled.
mosquito X1 (25 nL – 1.2 µL or 0.5 – 5 µL) is an automated single channel liquid handler. It’s low dead volume (< 0.5 µL) is ideal for DNA/RNA normalisation. mosquito’s easy-to-use software calculates the required volumes of buffer and DNA/RNA in order to perform the normalisation process seamlessly.
dragonfly is a larger volume dispenser (0.5 µL and above), capable of dispensing up to 10 reagents into any SBS plate format. Each disposable pipette works simultaneously and independently to dispense into any well without any cross-contamination, enabling fast, precise and accurate reagent dispense. Using dragonfly, it only takes under 4 minutes to dispense 4 reagents into a 96-well plate. Using true-positive displacement technology, dragonfly provides the freedom to use any liquid type, regardless of viscosity, without any need for liquid classification.
The technology – true-positive displacement pipetting
Unlike traditional pipettes, the mosquito robot uses true-positive displacement liquid handling technology. This superior pipetting mechanism allows dispensing in the 25 nL – 5 µL range with high precision and accuracy, across the volume range. Using this technology to prepare lower reaction volumes results in dramatic cost savings without compromising the complexity of the data, when compared to standard reaction volumes.
Miniaturisation of genomics reaction volumes is essential for a variety of applications such as PCR, qPCR , NGS sample preparation, Gateway® cloning, DNA quantification, gene expression analysis and synthetic biology. Lowering the reaction volumes provides:
- significant reduction of sequencing cost per base pair (reagent cost are a large proportion of sequencing costs)
- ultra-low input amounts of RNA or DNA (down to 20 pg or lower)
Furthermore, the use of disposable mosquito tips ensures zero cross-contamination and gentle sample handling.