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 from the cells or tissues using an RNeasy Plus Mini kit (Qiagen, Valencia, CA). One microgram total RNA and random primers were used for cDNA synthesis using the AffinityScript qPCR cDNA synthesis kit (Agilent Technologies, Santa Clara, CA). The PCR was performed using the following primers to identify the gBK/hbr5 region within the gBK transcripts: forward, 5′-CGTTGGGAAGAACATTGTTC-3′, reverse, 5′-AACTGGCTCGGTCACAAG-3′. After the thermal cycling was complete, the PCR products were run on a 1.5% agarose gel and stained with the ethidium bromide. The gel image was then captured under the UV light using the Eagle Eye II imager (Stratagene, San Diego, CA). The predicted size of the amplicons was 186 bp. The image was scanned for its densitometric reading by using the Un-Scan-It program (Silk Scientific, Orem, UT).

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Manufacturer protocol

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

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