The ability to amplify this quantity of RNA efficiently and effectively was one of our major design considerations.
The method also could provide significant merits for clinical applications with its ability to amplify lower quantity of samples and the cost-saving factors.
For example, library generation requires large quantities of DNA (µg) as starting material, and the use of this technology has thus been limited to studying cells or organisms where larger quantities of DNA can be readily isolated or where polymerase chain reaction (PCR) is used to amplify small quantities of DNA into sufficient quantities for library preparation.
Using the Polymerase Chain Reaction (PCR), which can specifically amplify large quantities of DNA from small starting quantities, sensitive identification procedures based on protozoan DNA sequence information can be developed.
PCR enables genetic material to be amplified in quantity both quickly and reliably.
In forensic science, particularly the field of DNA typing, this method is also applied due to its ability to amplify small quantities of DNA (Kashyap et al. 2004).
The RPM-Flu assay leverages a multiplexed RT-PCR-like strategy to rapidly (∼1 hour) amplify trace quantities of specific target gene sequences so that they may be detected if actually present in a specimen.
Multiple displacement amplification (MDA) with bacteriophage Phi29 DNA polymerase is emerging as the preferred method for WGA, as it can amplify minute quantities of DNA by several orders of magnitude with very high fidelity, producing fragments that are tens of kilobases long [2].
NuGEN discussed their Ovation® FFPE RNA Amplification System v2, which can be used to amplify picogram quantities of starting RNA, using a proprietary SPIA process for linear DNA amplification at constant temperature.
PCR based methods have been successfully used to amplify subnanogram quantities of RNA from as little as a single cell [ 13, 14], but these approaches have not been widely adopted.
The Ovation RNA-Seq V2 kit is able to amplify small quantities of RNA from tissue samples while maintaining transcript levels nearly identical to those from matched unamplified input RNA, demonstrating an overlap of 93% when comparing each matched pair individually.
