Highlights
- Universal Depletion: Novel probe-free technology depletes rRNA from any organism.
- Simplest Kit: Prepare NGS-grade RiboFree cDNA from total RNA in as little as 1.75 hours.
- Automation Friendly: Streamlined protocol for increased scalability.
Description
The Zymo-Seq RiboFree Universal cDNA Kit is the simplest kit for the preparation of NGS-grade first-strand cDNA with ribosomal RNA (rRNA) depleted. This simple cDNA kit features RiboFree Universal Depletion, a novel probe-free rRNA depletion technology that depletes rRNA from any organism. The RiboFree Universal Depletion is validated across biological kingdom and phyla, including mammals, plants, and various prokaryotes, and also RNA from a wide range of sample types including cells, snap-frozen tissues, and FFPE samples. The RiboFree single-stranded cDNA produced by the kit can be used for qPCR detection, RNA-Seq library prep, and any other downstream analysis. In contrast to poly(A) enrichment methods, the RiboFree Universal cDNA Kit produces single-stranded cDNA from nearly all RNA biotypes, including full-length mRNA, mRNA with degraded poly(A) tails, long-noncoding RNA (lncRNA), intronic RNA, nucleolar RNA, etc. The cost-effective Zymo-Seq RiboFree Universal cDNA Kit has been optimized to reduce hands-on time, with all the reagents such as SPRI beads included for researchers’ convenience.
Table 2. Zymo-Seq RiboFree Universal cDNA Kit (Cat. No. R3001) update summary
| | Current Version | Updated Version |
Instruction Manual Versions | v1.1.0 | v2.0.0 |
Total RNA Input Range | 100 ng – 5 µg | 10 ng – 1000 ng |
Depletion Reaction Duration* | 100 ng input – 4 hours | · 100 ng input – 1 hour
· 10 ng input – 4 hours |
* May vary depending on the quantity, quality, and source of the input RNA.
Equipment Needed (user provided) | Thermal cycler with heated lid, magnetic stand for 0.2 mL PCR tubes, microcentrifuge for 0.2 mL PCR tubes and 1.5 mL microcentrifuge tubes, and a benchtop vortex mixer. |
Sample Input Material | RNA |
RNA Input | 10 – 1000 ng of total RNA. - For optimal results, please use the recommended 10-1000 ng input.
- If an input below 10 ng is necessary, see Appendix B of the kit protocol for additional considerations and recommended modifications.
|
Input Quality | RNA should be free of DNA contamination and enzymatic inhibitors, with A260/A280 and A260/A230 ≥ 1.8. RNA with lower purity ratios (A260/A280 and A260/A230) should be treated with DNase I and purified with the RNA Clean & Concentrator™ (Cat. No. R1013) prior to processing. RNA should be suspended in water, TE, or a low-salt buffer. - For optimal results, please use intact RNA (RNA Integrity Number or RIN ≥ 8.0) whenever possible.
- For degraded RNA input, see Appendix C of the protocol for additional considerations and recommended modifications.
|
Processing Time | As little as 1.75 hours |