By using multiple probes against a single pathogen coupled with the proper amplification scheme, microarrays can serve as valuable tools for viral discovery, detection, genotyping and sequencing [13], [14], [21], [25], [26], [27], [28], [29], [30], [31], [32], [33].
This dissertation presents the design of an efficient class-AB amplification scheme based on a pseudo-differential, single-stage and cascode-free architecture.
Our results show a novel all-optical amplification scheme that relies on soliton induced scattering.
However, single amplification by enzyme labels is not sufficient for detecting an ultra-low analyte concentration.
The two-stage charge-to-voltage amplification scheme includes a numerical block for offset compensation.
The amplification protocol was optimized to ensure that a single amplification product was obtained.
If a single amplification product was obtained, the PCR product was sequenced directly using the amplification primers.
(right) Cartoon representation of PCR amplification scheme, with exon numbering according to zebrafish transcript.
We used the Applied Biosystem 7000 sequence detection system according to the recommended amplification scheme: 95°C 2 min and 40 cycles: 95°C 15 sec, 60°C 1 min. The dissociation curve method was applied according to the manufacturer's protocol (60°C to 95°C) to unsure the presence of a single specific PCR product.
Approximately 100,000-fold amplification is typical for a single amplification step.
