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 mosquito liquid handlers, have revolutionised sample prep workflows in many areas, including cancer and stem cell research, through high-throughput single cell analysis, using DNA or RNA sequencing. The following applications can benefit from using our versatile mosquito liquid handlers:
- Synthetic & molecular biology – High-throughput, miniaturised reactions for PCR, cloning and DNA assembly with mosquito.
- Nucleic acid sequencing – Low-cost, low-volume library preparation for single cell analysis, including DNAseq, RNAseq and normalisation.
- Gene expression profiling – Cost-effective, high-throughput analysis of mRNA with rapid qPCR set-up.
TTP Labtech’s mosquito liquid handling robots (single, 8- or 16-channel, volume range 25 -1200 nL or 0.5 – 5 µL) can be used for miniaturising a wide range of genomics applications. This unique true-positive displacement technology enables accurate low-volume library prep workflows validated with market leading kits including Illumina Nextera XT. At final volumes of 5 μL or less, miniaturised workflows reduce sample preparation costs by 90% or more, while providing increased throughput, process control and standardisation.
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 (25 nL – 1.2 µL or 0.5 – 5 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, standardised 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 and synthetic biology applications.
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.
Furthermore, the use of disposable mosquito tips ensures zero cross-contamination and gentle sample handling. Additionally, it offers the ability to pipette liquids with high viscosities (such as enzymes in 50% glycerol) accurately and precisely. It provides a unique opportunity to obtain the current level of data at a significantly lower cost, and/or to carry out larger scale studies at current costs.
Validated applications from major genomics collaborators include:
- Library preparation for single cell DNA and RNA sequencing.
- Reverse transcription using the SMARTer Ultra kit.
- Automated precise and accurate low-volume DNA normalisation.
- PCR and qPCR reaction set-ups.
- Gateway cloning.
- Colony spotting.
Featured in JALA with Louise Laurent’s lab (UCSD): Miniaturization Technologies for Efficient Single-Cell Library Preparation for Next-Generation Sequencing publication.
Different stages of human pancreatic stem cells were studies using single cell RNA-seq (scRNA-seq). 2, 4 and 8 uL Nextera XT libraries were prepared for technical and biological replicates and using mosquito. The libraries were also cleaned up in 384 format on mosquito. The data confirms that miniaturization of reaction volumes does not affect the scRNA-seq data quality. >> Read publication!
Featured in Cell by Irving L. Weissman’s lab (Stanford Medicine): Mapping the Pairwise Choices Leading from Pluripotency to Human Bone, Heart, and Other Mesoderm Cell Types.
As previously shown in JALA paper, miniaturisation of reaction volumes does not affect the scRNA-seq data quality. Here high throughput scRNA-seq was used to systematically catalogue the diversity of intermediate cell states formed during differentiation, and the minimal combinations of positive and negative signals that were sufficient for differentiation between each of these intermediate states was tested. mosquito X1 was used for the normalisation of cDNA concentrations, and mosquito HTS was used for Nextera XT library prep at 4 uL.
>> Read publication!
- Automated low-volume DNA normalisation and NGS library prep for single cell analysis (Prof Stephen Quake and Prof Irving Weissman lab, Stanford)
- Miniaturised Gateway cloning, PCR amplification and Nextera library generation (Novartis)
- Low-volume, single-cell RNA-seq library pre using Nextera XT (UC San Diego)