miRCURY LNA RNA Spike-in kit

For quality control of the RNA isolation and cDNA synthesis steps of miRCURY LNA miRNA PCR experiments

Features
•An easy and convenient control of your RNA samples and RT reactions
•Save time and reagents by identifying poor-quality samples early
•Available in an easy-to-use, pre-mixed format
•Highly flexible – use only the spike-ins you need for your study

Product Details

The RNA Spike-In Kit enables quality control of the RNA isolation, cDNA synthesis and PCR amplification
steps of miRCURY LNA miRNA PCR or miRCURY LNA miRNA Probe PCR experiments. Identifying any poor-quality
samples early in the process saves you time and reagents. Plus, the kit is easy to use and comes in a
premixed format that allows you to use only the spike-ins you need for your analyses.

Performance

Checking sample quality and PCR efficiency
Together with the miRCURY LNA miRNA QC PCR Panel, the RNA Spike-in Kit for RT offers a unique and comprehensive
 way of checking sample quality and PCR efficiency. The panel is compatible with many sample types, including
cell cultures, tissue samples and biofluids from human, mouse, rat, monkey and dog (see figure miRCURY LNA miRNA
 QC PCR Panels can be used with a wide range of sample types).

Test your isolated RNA to find outliers and suboptimal samples, including serum/plasma samples affected by hemolysis .
This saves time and resources in the laboratory.

The RNA Spike-In Kit can also be used for experiments using the individual assays with the miRCURY LNA miRNA PCR (for SYBR® Green) or miRCURY LNA miRNA Probe PCR systems.
 Individual assays for the relevant spike-in template are available in both the SYBR® Green and the probe-based miRCURY LNA PCR systems.

Principle

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.

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.

Procedure

Analysis and interpretation of data
An important factor when using the RNA Spike-In Kit is knowing how to interpret the experimental data. We have made this easy by devoting an entire section in the kit
handbook to explaining how to analyze and interpret the data in an easy-to-understand and clear-cut way. Refer to the RNA Spike-In Kit Handbook for details.

Applications

Quality control of the RNA isolation and cDNA synthesis steps of miRCURY LNA miRNA PCR and miRCURY LNA miRNA Probe PCR experiments