About YeaStar™ RNA Kit
The YeaStar™ RNA Kit provides all the necessary reagents for RNA isolation from a broad spectrum of fungi including: Aspergillus fumigatus, Aspergillus nidulans, Aspergillus nivens var. aureus, Candida albicans, Pichia pastoris, Saccharomyces cerevisiae, Schizosaccharomyces pombe. Generally, the kit can be used for the purification of high-quality, total RNA from any fungus that can be lysed by yeast lytic enzyme. The kit facilitates the purification of 10-25 µg of total RNA from 1-1.5 ml of cultured cells using innovative Fast-Spin column technology.
|Processing Time||30 min|
|Sample Sources||Fungi susceptible to yeast lytic enzyme lysis.|
|RNA Purity||High quality RNA is recovered in the supplied RNase-free water.|
|RNA Recovery||Typically, RNA is eluted into 60 µl RNase-free water. The provided columns have an RNA binding capacity of 25 µg.|
|Sample Size||1 to 1.5 ml liquid culture.|
Pellet 1-5 x107 cells (1-1.5 ml culture) by centrifugation at 500 x g for 2 minutes. Carefully remove all of the supernatant.
Add 80 µl of the YR Digestion Buffer and 5 µl of the Zymolyase to the cell pellet and resuspend the pellet completely by pipetting. Incubate the suspension at 30-37°C for 40-60 minutes2.
Add 160 µl of the YR Lysis Buffer and mix thoroughly by vortexing.
Centrifuge the mixture at 7,000 x g for 2 minutes3.
Transfer the supernatant to the Zymo-Spin™ IIIC Column in a Collection Tube and centrifuge at ³10,000 x g for 1 minute.
Add 200 µl RNA Wash Buffer to the column and centrifuge at ³10,000 x g for 1 minute. Discard the flow-through. Repeat the wash step.
Remove the Zymo-Spin IIIC Column carefully from the Collection Tube and transfer it into an RNase-Free Tube. Add 60 µl of DNase/RNase-Free Water directly to the column matrix4. Centrifuge at ≥10,000 x g for 30 seconds. The eluted RNA can be used immediately or stored at -70ºC.