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Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor

m quantities of DNA are amplified in all samples, eliminating the need for quantitation.

GenomiPhi DNA Amplification Kit representatively amplifies single- or double-stranded linear genomic DNA using Phi29 DNA polymerase to perform strand displacement amplification. Microgram quantities of representative DNA are produced from small amounts of starting material that can be used immediately in downstream applications.

For all kits, DNA amplification is achieved by combining the starting DNA with random hexamer primers, Phi29 DNA polymerase, dNTPs, and buffer components optimized for DNA synthesis. This reaction mixture is incubated overnight at 30 °C, during which time the available nucleotides are consumed and converted into high molecular weight fragment copies of the input DNA.


Automation
The amplification protocols are simple and allow straightforward automation. The Genesis Freedom series of RSPs from Tecan are ideal for automating DNA amplification reactions and downstream applications. The arm of the Tecan liquid handler is equipped with four or eight tips and can perform all required liquid handling steps with high sensitivity and accuracy. When the amplification reaction stops because the nucleotides in the reaction are exhausted, DNA yields are equivalent across all samples (Fig 2). Disposable tips eliminate cross-contamination concerns. With an eight-channel liquid handling arm, multiple 96-well plates of TempliPhi or GenomiPhi DNA amplification reactions can be prepared in half the time of manual pipetting. Whatever the post-amplification application (Fig 3), the open platform can be adapted. The Tecan Gemini software system allows easy protocol setup and operation of the workstation.



Conclusions
The combination of GE Healthcare’s DNA amplification products and
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