RNeasy Micro Kit

RNA isolation / purification Tissue - Mouse Cornea

Experiment
RNA isolation / purification Tissue - Mouse Cornea
Product
RNeasy Micro Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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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Discussion

Discussion

2 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

3 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Mouse Cornea using RNeasy Micro Kit from Qiagen.

Paper title
Treatment of donor corneal tissue with immunomodulatory cytokines: a novel strategy to promote graft survival in high-risk corneal transplantation
Full paper
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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