The soil I am working with comes from the Kalahari Desert that is a large semi-arid sandy savannah in Southern Africa. It is a sandy type soil with a high silica content.
I have successfully extracted RNA from a positive control (soil from a fungal pathogenicity trial) using the Qiagen RNeasy PowerSoil total RNA kit as well as the ZumoBIOMICS RNA Miniprep kit.
The sampled soils were snap freezed in liquid nitrogen using 3 methods: 1.) soil only, 2.) RNA Later and 3.) DNA/RNA Shield (Zymo).
I have tried to increase the sample input to up to 6g using the Qiagen (midi prep) kit and extracted DNA but no RNA.
Any suggestions would be appreciated.
I have successfully extracted RNA from the soil mentioned using the Qiagen RNeasy PowerSoil total RNA kit. Combining the beads and buffers (5 ml Bead sol, 5 ml Phenol mix, 1 ml of SRI & IRS) used for two extractions into a 50 ml capped tube (sterile) you can extract from up to 10 g of soil. Follow the normal protocol but increase the volume of SR3 at step 7 to 3 ml (not 1.5 ml). I also repeated the final incubation and centrifugation step (without inverting or mixing) and then I resuspend the final pellet in 50 uL of water. DNA contamination was present but can be eliminated using a DNase treatment.
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