DNAble Instrument Compatibility & Throughput
DNAble is rapid nucleic acid amplification system comprised of: a strand displacing polymerase, a nicking enzyme, target specific primers, target specific molecular beacon(s), and dNTPs. These components exponentially amplify and specifically detect target DNA sequences at a constant incubation temperature. Since there is no thermocycling required, DNAble provides results in as little as 5 minutes with high analytical sensitivity. Given DNAble’s unique amplification system, not all thermocyclers are recommended for use with DNAble. Only real-time thermocyclers and portable readers validated by EnviroLogix (such as the AmpliFire or T16 reader) are recommended for use with DNAble because the amplification system is so sensitive that real time fluorescence detection is required to accurately analyze results. When using a real-time thermocycler for DNAble, the activation, denaturation, and annealing stages are all set to 56 °C to achieve a constant incubation temperature. End point thermocyclers (e.g., hydrocyclers) are not recommended for use with DNAble because a baseline fluorescence reading cannot easily be obtained which leads to incorrect result interpretation in some instances.
To provide the industry access to DNAble’s testing benefits (crude sample preparation and fast time to results), EnviroLogix has validated DNAble chemistry on various liquid handling instruments and real time thermocyclers/readers. Instruments selected for validation were chosen to support a wide variety of use applications ranging from portable, low throughput applications to laboratory based, high throughput applications. Figure 1 depicts validated liquid dispensing instruments in relation to the testing throughput they support. Hand pipettes are sufficient for running low throughput assays in 8 well tube-strip format or running several 96 to 384 well plates a day. Liquid handlers are recommended for labs running >1,000 assays per day and are also utilized to automate sample preparation. For the highest throughput application, LGC’s IntelliQube is an all-in-one solution that automates sample and reagent dispensing, array tape sealing, assay incubation, and real-time detection.
- Hand pipette:
- single or multichannel: dispense volumes range from 1 to 25 µL
- Liquid handler:
- EpMotion 5073, dispense volume of 2.5 µL
- Tecan EVO, dispense volume of 2.5 µL
- Automated, fully integrated system:
- IntelliQube, dispense volume of 0.8 µL
When using hand pipettes or liquid handling stations for sample preparation and assay setup, a real-time thermocycler/reader is recommended to incubate the DNAble reaction and to detect fluorescence signal. Table 1 lists instruments that have been validated as compatible with DNAble.
Table 1: Instruments for DNAble Reaction Incubation and Real Time Fluorescence Detection
Values for throughput are wells analyzed per hour (includes sample preparation and amplification time)
|Instrument||# of Wells||Throughput||Instrument Description|
|Axxin T8||8||24 - 40||Semi - Portable|
|AmpliFire||8||24 - 40||Portable|
|Axxin T16||16||48 - 80||Semi - Portable|
|MyGo Pro||32||96 - 160||Semi - Portable|
|BioRad iQ2||96||288 - 480||RT thermocycler - Laboratory|
|Agilent AriaMx||96||288 - 480||RT thermocycler - Laboratory|
|Roche LC480||384||1,152 - 1,920||RT thermocycler - Laboratory|
|ViiA7||384||1,152 - 1,920||RT thermocycler - Laboratory|
|IntelliQube||768||1,536 - 2,126||Integrated Liquid Handling & RT thermocycler|