RNeasy Mini Kit From Qiagen

The isolation of total RNA from tissue samples is generally a starting point for other analyses and collection of data, such as real time RT-PCR, cDNA synthesis or microarrays. However, extraction of RNA is a process that must be performed stringently and be free of contamination. Samples may be easily contaminated with substances that break down the RNA present in a sample, in particular, ribonucleases (RNases), which are present on practically every surface and solution within a laboratory. This kit allows for purification of total RNA from a variety of sources, such as animal tissues, animal cells and bacteria as well as clean-up of RNA from other preparations, and the solutions provided are RNase free, which allows for extraction of RNA without degradation.

The RNeasy Mini Kit allows isolation of RNA from very small amounts of starting material and will yield up to 100 ug of total RNA. This RNA can then be quantified and used in other applications, such as RT-PCR, real time RT-PCR or microarrays. Isolation of RNA can be performed in less than 15 minutes, which is a significantly shorter period of time than traditional methods of extraction and is more environmentally friendly and less hazardous as it does not use phenol and/or chloroform extraction. The principle behind the methodology outlined in this kit involves a silica-based membrane and high-salt buffer system that allows extraction of up to 100 ug of RNA greater than 200 nucleotides in length.

In our laboratory, use of this kit has enabled us to extract RNA of high purity and substantial quantity from porcine animal mucosal tissue. When using animal tissue, it is first necessary to homogenize samples in the provided stabilization reagent to minimize any changes in gene expression patterns that may occur and eliminate RNases. After extraction and quantification, our RNA samples were then subsequently quantified and used in RT-PCR and real time RT-PCR analyses to determine expression levels of peptide transporters in intestinal tissue. Usage of this kit ensured that we could extract RNA and complete real-time PCR experiments in just one day.

I recommend this kit because it was very easy to use when compared to other RNA extraction kits on the market. The kit is very straightforward, with only a few additional materials needed, such as equipment for sample disruption and homogenization, beta-mercaptoethanol and a microcentrifuge, all of which are generally present in most laboratories. We have used this kit for isolation of RNA from animal tissues and we have analyzed samples for DNA contamination as well. The RNA we have isolated with this kit has been free of any contamination. It can be used for a wide variety of samples such as yeast, fungi, animal cells and plant cells. There is also a valuable troubleshooting section located in the product manual. We plan to continue using this kit in all of our studies involving real-time RT-PCR where high-quality RNA is essential.