RNeasy Midi Kit

RNA isolation / purification Tissue - Mouse ileum

RNA isolation / purification Tissue - Mouse ileum
RNeasy Midi Kit from Qiagen

Protocol tips

Upstream tips
- This kit purifies RNA from up to 20–250 mg tissue
Downstream tips
- If there is any DNA contamination in downstream applications incubate the RNeasy column for 5 min at room temperature with Buffer RW1 before centrifuging

Publication protocol

Total RNA was isolated from murine ileum and liver (RNeasy Midi Kit, Qiagen, Hilden, Germany). RNA integrity was tested by RNA gel electrophoresis. cDNA was synthesized from total RNA using the iScript cDNA synthesis kit (BioRad, Hercules, CA, USA). Quantitative RT-PCR was performed using SYBR Green Supermix (BioRad) on an iCycler iQ system using diluted cDNA as template (primer sequences are provided in Table S1 in File S1). Expression levels were estimated using the comparative threshold cycle method. Cyclophilin was used as housekeeping gene, with similar expression levels in ileum and liver under all experimental conditions.

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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?


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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Manufacturer protocol

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

Download PDF Download manufacturer protocol


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