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

The MARC-145 cell culture supernatant was used for RNA extraction with the help of RNeasy Plus Mini Kit (Qiagen,Mississauga, ON, Canada). 100 ng of RNA was added for RT-PCR using the Super Script™ III Platinum® Two-Step RT-qPCR Kit (Invitrogen). The cDNA, primer and IQ SYBR Green Supermix (Bio-Rad, Hercules, CA) were mixed together. The qPCR conditions were: 95°C for 3 min, followed by 45 cycles at 95°C each lasting 15 s (denaturation), then 3 s at 55°C (annealing), and finally 30 s at 72°C (elongation). The normalized amounts of PRRSV genome relative to the VLP vaccination group were analyzed by the 2-ᇞᇞCt method (Schmittgen and Livak, 2008).

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Manufacturer protocol

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

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