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 isolated with the RNeasy Plus Mini Kit (QIAGEN). RNA was reverse transcribed into cDNA using the iScript cDNA kit (Biorad), and the cDNA was used for Quantitative PCR (SYBR green qPCR kit, Biorad). Each qPCR reaction was run in triplicate and was normalized to the GAPDH gene transcript. Gene expression was quantified using the 2−ΔΔCT method (mean fold change of gene expression = 2−ΔΔCT). The primers used for real-time PCR analysis of IDO were as follows: sense: ACT GTG TCC TGG CAA ACT GGA AG, antisense: AAG CTG CGA TTT CCA CCA ATA GAG.

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Manufacturer protocol

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

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