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Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000

r automation in mind. It is possible that the two kits could be fully automated to run sequentially. However, storage and transport solutions would need to be considered for this system integration. This is another aspect of "scalability", in that the semi-automated system presented here could be integrated into a fully automated system. The MJ Research Hard-ShellThin Wall plates were chosen for their rigidity before and after thermocycling. Plates that maintain constant physical dimensions are crucial for automation.

Travel entry and exit speeds of 20% and 5% were chosen based on observations where rapid entry and exit can cause non-uniform droplet breakoff. Higher entry speeds would likely have little affect, however, exit speeds can have a dramatic affect on precision and accuracy, as evidenced by visual observation of droplet residue left on the outside of pipetting devices.

Tips are ejected immediately prior to loading in our protocols as a matter of programming style. This style takes into account the case where a user has paused or stopped a method with tips left on.

The precision and accuracy of the Sciclone ALH 3000 was sufficient to produce yields comparable to the manual results. Since the well-to-well %CV was <4.2 and the smallest volume transfer was 0.5 μL, it is reasonable to infer that the Sciclone ALH 3000 performed within its operating specifications for the types of liquids used in these kits, that the PCR reaction provides a leveling effect, or both.

Conclusions:

Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000, produce data equivalent or superior to historical manual results3. Since all of this work was performed on an "out-of-the-box" system, it is likely that optimization of the system and liquid classes
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