RNeasy Micro Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.		- Always blank the spectrophotometer with your elution buffer. -When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Upstream tips
This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.
Downstream tips
- Always blank the spectrophotometer with your elution buffer. -When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Publication protocol

Real-time PCR
Using Trizol (Ambion, Life Technologies, Grand Island, NY) tissues were homogenized on ice and RNA was precipitated in the aqueous phase using 70% ethanol, followed by subsequent extraction and purification using RNeasy Micro kit (Qiagen, Valencia, CA, USA). Reverse transcription of total RNA was conducted using the Superscript III kit (Invitrogen, Carlsbad, CA, USA). Real-time PCR was performed using TaqMan Universal PCR Mastermix (Applied Biosystems, Foster City, CA, USA) and preformulated primers for IL-10 (Mm00439614_m1), TNF-α (Mm00443258_m1) and GAPDH (Mm99999915_gl). The PCR conditions were 40 cycles at 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, followed by final extension at 72 °C for 10 min. The results were analyzed by the comparative threshold cycle method and normalized to GAPDH as an internal control.

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Manufacturer protocol

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

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