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

Spleens were collected in RNA later (Ambion, Austin, TX, USA) and RNA isolated using RNeasy Mini kit following the manufacturer’s protocol (QIAGEN, Valencia, CA, USA). RNA (500 ng each) was reverse transcribed to cDNA using Superscript VILO cDNA Synthesis kit (Invitrogen Corporation, 11754–250, California, USA) and Real Time PCR was performed using SYBR Green Core Reagent kit and a 7300 RT-PCR system (Applied Biosystems, Austin, TX, USA). Primers are listed in S2 Table. GAPDH was used for normalization.

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Manufacturer protocol

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

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