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

Total RNA was extracted from RPASMCs using RNeasy mini kit (Qiagen, Valencia, CA) and quantitative RT-PCR (qRT-PCR) were performed on the cDNAs generated from 250 ng of total RNA by HotStart-IT SYBR Green qPCR Master Mix with UDG (2×) Tested User Friendly kit (USB Corporation, Santa Clara, CA). A complete list of primers used in this study is provided in Table E1 in the online supplement.

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Manufacturer protocol

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

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