RNA isolation tips
Successful isolation of high-quality RNA from soil
RNA isolation from soil is one of the most difficult applications performed in environmental molecular biology. Not only is degradation a constant challenge, but also RNA yields tend to be low.
Possible co-contamination of the RNA by soil humic acid and inhibitor content is also a challenge. Purity for RNA applications is essential. The RNA should be concentrated when added to the reaction and inhibitors cannot be present.
Low RNA yields, usually between 10 and 20% of the yield of DNA, mean that to isolate a sample for research requires working with a larger amount of sample, using larger (15ml) tubes and a centrifuge with a greater capacity. As dilution of the RNA for reverse transcription is best avoided when looking for low copy genes, successful inhibitor removal is key.
To meet these challenges, QIAGEN® offers the RNeasy® PowerSoil® Total RNA Kit, which has been specifically developed to make soil RNA extraction straightforward.
The protocol is a carefully-tested combination of methods: Inhibitor Removal Technology (IRT®) for inhibitor removal, phenol-chloroform extraction for complete microbial lysis, and anion-exchange for high-quality purification. The end result is the isolation of clean RNA in the volume desired, allowing for maximal use in RT-PCR.
From loamy forest soil to river sediment, every soil has a different texture, moisture and microbial load. Using the RNeasy PowerSoil Total RNA Kit, you can adapt your extraction process for each one.