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RNA Spike-In Kit

RNA spike-in kit for quality control of the RNA isolation and cDNA synthesis steps of a miRCURY LNA™ Universal RT microRNA PCR experiment.

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

Features

Use the RNA spike-in kit for quality control of the RNA isolation and cDNA synthesis steps of a miRCURY LNA™ Universal RT microRNA PCR experiment. 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, important information regarding the quality of the samples can be obtained (Table 1).


Table 1. Overview of issues and conclusions


 

Product coverage

Exiqon’s RNA spike-ins (UniSp2, UniSp4, UniSp5, UniSp6) resemble microRNAs in structure but lack close sequence similarities to known microRNAs. In addition, the kit includes a synthetic version of the C. elegans microRNA, 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 100 fold increments (Table 2). This set of spike-ins is intended as an RNA isolation control.
  • Vial 2: Contains a synthetic version of cel-miR-39-3p. This microRNA is meant to be used in conjunction with the UniSp6 spike-in provided with the Universal cDNA Synthesis Kit (denoted “RNA spike-in”). Mixing of these RNA templates as described in the manual will result in a 100 fold concentration difference between the RNAs (Table 2). This set of spike-ins is intended as a cDNA synthesis control.
Each vial contains RNA sufficient for 50 reactions. In addition, they contain MS2 carrier RNA intended to improve the stability of the RNA spike-ins without interfering with down-stream reactions.

Table 2. Contents of the RNA Spike-In Kit



Choose the spike-in RNAs you prefer

In order to take 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 Exiqon’s SYBR® Green Master Mix). This means that you have the flexibility to use only the spike-ins you need and don’t have to pay extra for primer sets you are not going to use.


Compatibility

The RNA Spike-in Kit is compatible with Exiqon Pick-&-Mix microRNA PCR Panels and individual miRCURY LNA™ Universal RT microRNA PCR assays.




Analysis and interpretation of data

An important factor when using the RNA Spike-In Kit is knowing how to interpret the experimental data. Exiqon has made this easy by devoting an entire section in the manual to explaining how to analyze and interpret the data. Learn how to analyze your data in an easy-to-understand clear-cut way: Download the RNA Spike-In manual


Figure 1 The RNA Spike-in kit was used for qPCR quality control of commercially available plasma from two different sources (n=2 for each). UniSp2 / UniSp4 / UniSP5 RNA Spike-in mix was introduced in the RNA isolation step (blue) and the UniSp6 / cel-miR-39-3p RNA Spike-in mix was added to the RT reaction (red) as described in the Instruction Manual. As seen from the Cq values, the kit offers a wide dynamic range which is ideal for control of the RNA isolation and RT reaction steps, respectively. Moreover, the figure shows that the RNA isolation and the RT reaction were highly reproducible.
Exiqon’s RNA Spike-In Kit covers the full range of microRNA expression levels. (Click to learn more)


NOTICE TO PURCHASER: LIMITED LICENSE Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser’s own internal research. No other patent rights are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. SYBR® Green is a trademark of Invitrogen.
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