miRNeasy Mini kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.

Protocol tips
- To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.

Publication protocol

Total RNA was extracted from the moDCs, cultured in the presence of the indicated stimuli 6 h or 24 h post stimulation, for messenger RNA (mRNA) or microRNA (miRNA) analysis respectively. The Phenol extraction column-based miRNeasy RNA isolation kit (Qiagen, Venlo, the Netherlands) was used. Murine lung tissue collected on day 3 and day 7 after infection was homogenized by using the FastPrep®-24 bead homogenizer (MP Biomedicals, Santa Ana, CA, USA) in 700 µL Qiazol (Qiagen). RNA was isolated according to the manufacturer’s instructions with minor modifications, namely an extra column-washing step was added using 80% ethanol before extracting the RNA using RNase-free water. RNA quality and integrity was determined using Lab-on-Chip analysis on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The RNA integrity numbers (RIN) of all RNA samples were >8.



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Manufacturer protocol

Download the product protocol from Qiagen for miRNeasy Mini kit below.

Download manufacturer protocol

Videos

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