RNeasy Lipid Tissue Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- - If the RNA yield is low, repeat RNA elution, but incubate the RNeasy Mini spin column on the benchtop for 10 min with RNase-free water before centrifuging. - All centrifugation steps should be performed at 15–25°C except for phase separation

Protocol tips
- - If the RNA yield is low, repeat RNA elution, but incubate the RNeasy Mini spin column on the benchtop for 10 min with RNase-free water before centrifuging. - All centrifugation steps should be performed at 15–25°C except for phase separation

Publication protocol

Perfused epididymal adipose tissues harvested from LPS-injected mice were cut into 30 mg (approximate size) pieces and digested with 1 mg/mL collagenase (Type I, Sigma-Aldrich, St. Louis, MO, USA) dissolved in 2% BSA-HBSS + 5mM glucose for 1h with agitation at 37°C. Digested cells were then passed through a 100μm cell strainer and the strainer was washed with HBSS, followed by centrifugation for 10 minutes. Floating adipocytes and buffer were transferred to a new tube, followed by a second centrifugation. The pellet containing the SVF was resuspended in HBSS and transferred to a new tube, followed by centrifugation. All centrifugation steps were performed at 4°C and 450 × g. Total RNA was isolated using RNeasy® Lipid Tissue Mini Kit (Qiagen, Valenica, CA, USA).

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Manufacturer protocol

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

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