RNA Amplification
While most microarray experiments can now be completed with less than half a microgram of total RNA, there are still samples for which that amount is impossible to obtain. In these cases, we amplify the RNA using a kit from NuGEN, Inc. NuGEN kits provide a consistent, whole transcript amplification that introduces minimal bias.
| Kit | Application | Concentration | Volume |
|---|---|---|---|
| WT-Ovation Pico | Small amounts of intact RNA | 6-25 ng/µL | 8 µL |
| WT-Ovation FFPE v2 | Degraded RNA (eg FFPE samples) | 20-75 ng/µL | 7 µL |
| WT-Ovation FFPE v2 | Very small amounts of intact RNA | 2.5-10 ng/µL | 7 µL |
- WT-Ovation Pico (WT-Pico): For most amplification projects, this is the kit that best balances price and performance. Total RNA input must be between 500 pg and 50 ng. Inputs above 50 ng per reaction may inhibit amplification, while lower amounts of input will potentially result in insufficient yields depending on required analytical platforms.
- WT-Ovation FFPE v2 (FFPE): This kit is specifically intended for amplification of FFPE samples and other degraded RNA. However, it also works very well for small amounts of intact RNA. As samples move toward the lower limit of our detection capabilities, we prefer to use the FFPE kit for a more robust amplification. Total FFPE-derived RNA input must be between 50 to 100 ng. Inputs above 100 ng per reaction may inhibit amplification, while lower amounts of input under 50 ng may result in insufficient yields for analysis.
Both of these kits can be used for many microarray platforms, and can potentially amplify even smaller amounts than are listed here. We encourage investigators with questions about RNA amplification to contact us.