Features
Use the RNA Spike-In Kit for quality control of the RNA isolation and cDNA synthesis steps of your miRCURY LNA miRNA PCR or miRCURY LNA miRNA Probe PCR experiments. It is important for any qPCR experiment to ensure that the quality of the input RNA is sufficiently high for effective amplification. By introducing spike-in RNAs during RNA isolation and cDNA synthesis and comparing the qPCR results to those from endogenous reference genes, you can gain important information about the quality of your samples (see figure Overview of issues and conclusions). The kit offers a wide dynamic range, which is ideal for controlling these processes (see figure The RNA Spike-in Kit covers the full range of miRNA expression levels).>
Product coverage
Our RNA spike-ins (UniSp2, UniSp4, UniSp5, UniSp6) resemble miRNAs in structure but lack close sequence similarities to known miRNAs. The kit also includes a synthetic version of the C. elegans miRNA, cel-miR-39-3p.
The RNA Spike-In Kit is distributed in two vials:
- Vial 1 contains 3 RNA spike-ins (UniSp2, UniSp4 and UniSp5) pre-mixed, each at different concentration in 10-fold increments (see table below). This set of spike-ins is intended as an RNA isolation control.
- Vial 2 contains a synthetic version of cel-miR-39-3p. This miRNA is meant to be used in conjunction with the UniSp6 spike-in provided with the miRCURY LNA RT Kit (denoted "RNA spike-in"). Mixing these RNA templates as described in the kit handbook will result in a 100-fold concentration difference between the RNAs (see table below). This set of spike-ins is intended as a cDNA synthesis control.
Each vial contains RNA sufficient for 50 reactions. Also included is MS2 carrier RNA, which improves the stability of the RNA spike-ins without interfering with downstream reactions.>
Contents of the RNA Spike-In Kit | Reagent | Amount | Conc. (resuspended) |
UniSp2, UniSp4, UniSp5 RNA Spike-in template mix: | Synthetic UniSp2 RNA (22 nt) | 160 fmol | 2 fmole/µl |
| Synthetic UniSp4 RNA (22 nt) | 1.6 fmol | 0.02 fmole/µl |
| Synthetic UniSp5 RNA (22 nt) | 0.016 fmol | 0.00002 fmole/µl |
| MS2 total RNA | 50 ng | 0.625 ng/µl |
cel-miR-39-3p RNA Spike-in template: | Synthetic cel-miR-39-3p RNA | 0.16 fmol | 0.002 fmole/µl |
| MS2 total RNA | 50 ng | 0.625 ng/µl |
Choose the spike-in RNAs you prefer
To take full advantage of the RNA Spike-In Kit, you will need to purchase the corresponding LNA primer sets separately, with the exception of UniSp6, which is included in the miRCURY LNA RT Kit. This gives you the flexibility to use only the spike-ins you need without having to pay extra for primer sets you are not going to use. >
Compatibility
The RNA Spike-In Kit is compatible with the miRCURY LNA miRNA PCR Assays and Panels.