mosquito HV for Serial Dilutions
The TTP Labtech mosquito HV is a cost-saving assay miniaturisation solution for PCR and next generation sequencing setup.
The mosquito HV liquid handler bridges the gap between nanolitre and microlitre pipetting. It provides accurate liquid handling for low volume serial dilutions and assay plate preparation across the volume range of 500nL – 5µL.
Benefits For Screening & Molecular Biology
TTP Labtech’s mosquito HV liquid handler offers the following benefits:
- Serial dilutions can be carried out using cost-saving, miniaturised volumes in standard plates. Final volumes of between 2 – 8µL of serially diluted samples are easily achieved.
- Dilution plates can be stamped direct from assay plates, as part of the same protocol for additional time saving
- Reformat between different plate densities and types in the same protocol, accurately pipetting between 96, 384 and 1536 well plates.
- Able to create acoustic-ready source plates to maximise workflow efficiency
- Simple, fast, reduced volume PCR/qPCR plate set-up
- Miniaturized NGS library preparation, reducing cost
Benefits For Protein Crystallography
TTP Labtech’s mosquito HV also facilitates protein crystallography in the following ways:
- Ideal for the accurate creation of larger volume drops (1 to 10 µL) of multiple components used in scale-up and optimisation work
- Precise pipetting across a wide range of liquid viscosities with no format change required
- Zero cross-contamination of samples is ensured with TTP Labtech’s unique disposable micropipettes
The mosquito HV liquid handler can be used in applications including:
- Compound and small molecule screening
- Next generation sequencing library preparation
- molecular biology applications including rtPCR/qPCR set-up, RNAi screening
- SNP genotyping.
- Cell- or bead-based assays
- Enzyme kinetic assays
Percentage accuracy of target volume and repeatability (% c.v.) of DMSO obtained from 384 repeat volumes.
mosquito® – Automated Solution for Miniaturising NGS Sample Preparation
The introduction of automated nanolitre volume liquid handling instrumentation revolutionises cloning and NGS (Next Generation Sequencing) workflows, provided that the necessary accuracy and precision are maintained across the wide range of different liquid types used in these reactions. The miniaturisation of plate setup and reagent addition steps would facilitate high throughput applications, which in turn, offers excellent cost saving advantages over traditional large volume workflows.
Protocol for Miniaturising NGS Sample Preparation
We have been successful in miniaturising Nextera NGS sample preparation. The protocol involves four major steps:
- PCR amplification
- PCR cleanup
In our application note, we show the effect of reducing the total reaction volume from 50 µL (original protocol) down to 1 µL. At the lowest reaction volume (1 µL), 100 nL TDE1 and 1 ng (400 nL) of plasmid DNA were added using the mosquito – see the table below for further experiments:
|DNA (µL, ng)||20, 50||10, 25||4, 10||2, 5||0.4, 1|
|TD Buffer (µL)||25||12.5||5||2.5||0.5|
The mosquito easily handled the reagents and samples that were used in these reactions, down to 25 nL, with high accuracy and precision of 7 and 5% respectively. In this study, samples were sequenced at 2 x 50 paired end reads. Following sequence analysis, reactions showed 100% coverage for all 96 plasmids, with every nucleotide having at least 64 reads of coverage. The data clearly shows that miniaturising did not affect the data quality in any way.
Additional Benefits of Using the mosquito® for NGS Applications
TTP Labtech’s mosquito offers significant benefits for high throughput gene amplification and sequencing applications:
- Reduced cost through miniaturisation of sample prep
- Lower consumption of genomic DNA
- Gentle pipetting resulting in less shearing of DNA or frothing/denaturing of the enzyme
- Ability to pipette liquids with high viscosities (such as enzymes in 50% glycerol) accurately and precisely.