Miniaturized Genotyping Reactions Using the Echo Liquid Handler
The Echo Liquid Handler enables low volume genotyping reactions in a 384-well format with as little as 500 nL total reaction volume. Precise and accurate drop placement eliminates cross-contamination, while offering the flexibility to transfer from any well of a source plate to any well of a destination plate. Cost benefits from reduction of tip waste and decrease in reagent consumption can be realized without sacrificed data quality.
- Miniaturize PCR to save reagent cost and waste
- Increase throughput with lower volume 384-well PCR
- Contamination-free reagent and sample transfer
- Eliminate pipette tips and washing to save consumables cost and waste
Miniaturized Genotyping Reactions
| Components | Volume (nL/rxn) | Volume (nL/rxn) | Volume (nL/rxn) | Volume (nL/rxn) | Final Conc. | Final Rxn Conc. |
| Water | 712.5 | 536 | 356.25 | 78.125 | 1.5X | 1.125X |
| 5X DNA Probes Master | 450 | 337.5 | 225 | 112.5 | 1.5X | 1.125X |
| Primer Mix (20X) | 112.5 | 84.5 | 56.25 | 28.125 | 1.5X | 1.125X |
| Genotyping Probe, 10X | 225 | 167 | 112.5 | 56.25 | 1.5X | 1.125X |
| Total MM Volume Dispensed | 1500 | 1125 | 750 | 375 | 1.5X | 1.125X |
| DNA Plasmid Dispensed | ||||
| WT Plasmid | 500 | 375 | 250 | 125 |
| Heterozygote | 500 | 375 | 250 | 125 |
| Mutation | 500 | 375 | 250 | 125 |
| Total Reaction Volume | 2000 | 1500 | 1000 | 500 |
- Miniaturize PCR as low as 500 nL
- Simple assay setup allows for varying conditions within one microplate
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Echo 555 Liquid Handler |
Figure 1: (Right) Quantification of wild-type, heterozygote and mutant in 1.0 µL reaction volume |
Zero Cross-Contamination Results
Zero cross-contamination schematic for alternating WT, HET, and MUT plasmid plate.
Positive/Negative Call by the LightCycler 480. Red circles indicate positives. Green circles indicate negatives. There were no positive calls for mutant genes indicating a 100% call rate and zero cross-contamination.
Zero cross-contamination with multiple transfer steps
Reactions were dispensed in 2000 nL final reaction volumes in a two step process utilizing the Echo Liquid Handler. DNA Probes Master mix including the primer mix and genotyping probe was dispensed, followed by alternating plasmid DNA (wild-type (WT), heterozygote (HET), and mutation (MUT)) in an alternating pattern, shown above.
- Starting with only a few source wells, the Echo Liquid Handler can transfer from any well to any well with zero cross-contamination.
- Tipless transfer of reagents can occur without adherence to a format imposed by traditional liquid handlers.
- Zero cross-contamination with multiple transfer steps of primer, probe, mastermix and samples.
The Echo 500 Series Liquid Handlers revolutionize liquid transfer by using acoustic energy to eject fluids. The Echo Liquid Handler is completely touchless—no tips or nozzles, and no material contacts the same as it moves from source to destination. Low-volume, accurate and precise transfer at volumes of 2.5 nL and up allow for assay miniaturization to previously unattainable volumes. Acoustic transfer is compatible with a wide range of fluid types, enabling high-throughput total PCR assay assembly for applications including genotyping, gene expression and DNA sequencing. Transfer of samples from any well of a source to any destination further minimizes sample consumption and simplifies assay setup and execution.
