RNeasy Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- 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 and Proteinase-K treatment. - 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
- 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 and Proteinase-K treatment. - 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

For total RNA isolation, RNeasy Mini Kit (Qiagen, Hilden, Germany) was used and the isolation was made according to the manufacturer's instructions. Also RNA from CML cell lines K562, BV-173 and CML-T1 was isolated using the same kit. To remove possible DNA contamination, a DNase free set (Qiagen) was used as instructed by the manufacturer. cDNA was synthesized from up to 0,5 µg RNA with iScript cDNA synthesis kit according to the manufacturer's instructions (BioRad, Hercules, CA, USA).

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Manufacturer protocol

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

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