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. - 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. - 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 was extracted from J6/JFH-1-huh 7.5 cells or naïve huh 7.5 cells cocultured with NK-92 under various conditions using an RNeasy Mini kit (Qiagen, Illumina, CA, USA), and then synthesized into cDNA using M-MLV Reverse Transcriptase (BioTech, New York, NY, USA). The resulting cDNA was analyzed for the expressions of HCV genes by qRT-PCR. Gene expression levels were determined using SYBR Green reagent (Bioline, Trento, Italy) and a Step One Plus Real-time PCR system (Applied Biosystems, Foster City, CA, USA), while 18sRNA was used as an internal control. PCR primer sequences were as follows: J6/JFH1, J6/JFH1F (5′-CTTCACGCAGAAAGCGCCTA-3′)/J6/JFH1R (5′-CAAGCGCCCTATCAGGCAGT-3′).

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Manufacturer protocol

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

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