Extraction of RNA from yeast using the ZymoBIONICS YeaStar RNA kit
Quick, spin-column purification of total RNA (including small/microRNAs) from a wide range of fungi species, susceptible to yeast lytic enzyme lysis. Omits the use of mechanical homogenization and organic denaturants. RNA is ready for Next-Gen Sequencing, RT/qPCR and any downstream application, etc.
sample_prepVersion History
Version 1 Current
Effective: 2025-11-04First version.
Procedure Steps (Version 1)
Pellet 1-5 x 10^7 cells (1-1.5 ml culture) by centrifugation at 500 x g at room temperature for 2 minutes. Then carefully remove the supernatant.
Add 80 μl of YR Digestion Buffer and 5 μl of Zymolyase to the cell pellet and resuspend the pellet completely by pipetting up and down.
Incubate the suspension at 37 degrees C for 40-60 minutes. NOTE: If the volume of the cell pellet is ≤ 25 μl, incubate for 40 minutes. For cell pellet volume ≥ 25 μl, incubate for 60 minutes.
Add 160 μl of the YR Lysis Buffer and mix thoroughly by vortexing.
Add an equal volume ethanol (95-100%) (1:1) and mix well. Example: Add 245 μl ethanol to 245 μl mixture (sample in YR Lysis Buffer).
Transfer the mixture to the Zymo-Spin™ IIICG Column in a Collection Tube and centrifuge at 10,000 x g for 30 seconds. Discard the flow-through. (Optional) At this step, DNase I treatment can be performed with the DNase I Set and RNA Prep Buffer (#E1010 and #R1060-2-25; available separately).
Add 200 μl RNA Wash Buffer to the column and centrifuge at 10,000 x g for 30 seconds. Discard the flow-through.
Add 200 μl RNA Wash Buffer to the column and centrifuge. Then carefully, transfer the column into a nuclease-free tube (not provided).
Add 60 μl of DNase/RNase-Free Water directly to the column matrix and centrifuge at 10,000 x g for 30 seconds. Alternatively, for highly concentrated RNA use ≥ 35 μl elution. The eluted RNA can be used immediately or stored frozen.