Total RNA isolation with RNeasy kit (Qiagen)

Contributed by Dr. Alexei Gratchev

With RNeasy RNA isolation became really simple. You can lyse fresh or frozen cell pellet or even the cells directly on the plate (as it actually possible with most of existing RNA isolation system). RNA is then boud on the column, washed and eluted. The most problematic step is elution. You have to use relatively high volume of water, what resultes in the low concentration of RNA. If you use smaller volume the losses will increase dramatically. However the columns have a very good size cut off, and RNA preparation contains almost no tRNA. Another advantage of this kit usage is that all required RNase free reagents and materials are provided. The time you need for one preparation is about 2 h depending on the number of samples you have to process. You have to choose the right size of the preparation kits are avaliable in mini, midi and maxi format covering the range from 10e4 to 10e9 cells. This kits are a good choice for one who work with a big number of samples and need not a highly concentrated RNA. The manual is comprehensive and describes the very details of the protocol. More information can be obtained from Qiagen web site.

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