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 extracted from the sixteen cell lines harvested at log phase, using the RNeasy® Plus (Qiagen) kit following the manufacturer’s protocol. For real-time quantitative PCR (qPCR), mRNA was reverse-transcribed into cDNA using an ABI RT kit (Applied Biosystems). qPCR primers and probes were designed using Roche Probe Finder Design Software (Roche Applied Sciences) and reactions performed using the ABI PRISM 7900 Sequence Detection System instrument and software (Applied Biosystems). The reactions were incubated in a 384-well optical plate at 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 10 min. Experiments were performed in triplicate for each sample. Gene expression was normalized to internal control SDHA (succinate dehydrogenase), YWHAZ (tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide).

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Manufacturer protocol

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

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