RNeasy Plus Mini Kit

Protocol tips

Upstream tips	Protocol tips	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.

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

Total RNA was isolated with the RNeasy® plus mini kit (QIAGEN) according to the manufacturer's protocol. In the animal experiment, total RNA from livers was isolated with Tri-Reagent® (Sigma-Aldrich) and DNase treated with RapidOut DNA Removal Kit (Thermo Scientific) according to the manufacturer's protocols. Quantity and quality of the total RNA were determined by NanoDrop by analysis of the ratios at 260/280 nm and 260/230 nm.

In the cell work RT-PCR was performed with 10 ng of the isolated RNA in each reaction (total volume per reaction was 20 μL) with the QIAGEN® OneStep RT-PCR kit (QIAGEN) and following the manufacturer's protocol. In the animal experiments, RT-PCR was performed with 25 ng of cDNA in each reaction (total volume per reaction was 25 μL) with the HotStarTaq Plus DNA Polymerase kit (QIAGEN) and following the manufacturer's protocol

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Manufacturer protocol

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

Download manufacturer protocol

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