What methods do you recommend for RNA isolation?
We recommend a column-based method, including:
- Norgen Biotek Total RNA Purification Kit
- Zymo Research Quick-RNA™ Kits
- Arcturus PicoPure® RNA Isolation Kit
- Ambion PureLink® RNA Mini Kit
- Qiagen RNeasy Kits
Organic methods such as TRIzol® Reagent should be subsequently followed with a column-based clean-up method.
Can I use TRIzol® or other phenol-chloroform based extractions for RNA isolation?
We do not recommend the use of TRIzol or similar methods as any carryover of organic solvents may inhibit downstream enzyme activity. If using TRIzol extracted RNA, we recommend using a column-based purification of the RNA prior to input into the kit.
Can I use carrier RNA during RNA isolation?
We do not recommend the use of carriers during RNA isolation. If a carrier is required, please contact Tecan NGS Technical Support for more information.
How much total RNA do I need for amplification?
We recommend using between 500 pg and 50 ng total RNA as starting material. Input amounts outside this range may produce unsatisfactory and variable results.
Do I need to use high-quality total RNA?
RNA samples of high molecular weight with little or no evidence of degradation will amplify very well with this product. However, due to the whole transcriptome amplification approach, lower quality RNA samples and transcripts with a compromised poly(A) tail can also be amplified successfully using the Ovation Pico WTA System V2. However, the RNA should have high purity, and be free of contaminants.
Can contaminating genomic DNA interfere with the amplification performance?
Yes. This system is designed to amplify RNA, but contaminating genomic DNA may amplify during the process. For this reason we recommend DNase treatment during RNA purification.
Do you recommend DNase treatment of my total RNA sample?
Yes. For an explanation of DNase requirements see section III.A.4 of the User Guide. You may also find recommended procedures for DNase treatment in the Appendix.
Can I use the Ovation Pico WTA System V2 on bacterial RNA samples?
The Ovation Pico WTA System V2 amplification process has been shown to work with some bacterial RNAs. However, the kit has not been optimized for this purpose.
Are there any tissues that will not work with the Ovation Pico WTA System V2?
We have not encountered any specific RNA sources that will not work with the Ovation Pico WTA System V2. The RNA should have high purity and be free of contaminants.