RNeasy Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For Candida albicans, a more specific protocol might be required (see reference). To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	"- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C"

Protocol tips
For Candida albicans, a more specific protocol might be required (see reference). To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
"- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C"

Publication protocol

RNA extractions were performed using Qiagen RNeasy Mini Kit (cat#74104) with the following modifications. Cells were resuspended from the membranes with 2 x 900 μl ice cold water washes with vortexing for 30 sec after each addition. 1.5 mls of cell suspension was added to prechilled microfuge tubes and centrifuged at top speed for 30 sec. 600 μl RLT + 1%BMSH was added to resuspend the cells and this was added to a fresh 2 ml screw cap tube containing 300 μl Zirconia beads (Ambion, Fisher Scientific) and 600 μl phenol:chloroform:isoamyl alcohol 25:24:1, vortexed on a mini-beadbeater (Biospec Products) for 3 min, and centrifuged at 14000 rpm for 5 min at 4°C. 550 μl of the aqueous phase was transferred to a new microfuge tube, an equal volume of 70% ethanol added, and RNA isolation proceeded as the manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Mini Kit below.

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