RNeasy Mini Kit

RNA isolation / purification Tissue - Human ileum

Experiment

RNA isolation / purification Tissue - Human ileum

Product

RNeasy Mini Kit from Qiagen

Manufacturer

Qiagen

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C.

Protocol tips
- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C.

Publication protocol

RNA was purified from frozen tissue using the RNeasy Mini Kit (Qiagen, Copenhagen, Denmark) according to manufacturer's instructions. For each batch of purification, a maximum of 30 mg of tissue was homogenized at 25 Hz for 3 min using a Tissuelyser II machine (Qiagen) and 5 mm Stainless Steel Beads (Qiagen). Homogenized tissue was purified using RNeasy Mini columns, and subsequent to purification, on‐column DNase digestion of the RNA for elimination of DNA contamination was performed (RNase‐Free DNase Set, Qiagen). RNA was purified from FFPE samples/formalin‐fixed tissues using the RNeasy FFPE Kit (Qiagen). Five sections of 10 μm were pooled and deparaffinated in deparaffination Solution (Qiagen) and RNA was subsequently purified according to manufacturer's instructions for optimized removal of genomic DNA contamination. Quantification of total RNA was measured using a Thermo Scientific NanoDrop 1000™ (Thermo Scientific, Hvidovre, Denmark) and Thermo Scientific Software ND.1000 V3.5.2.

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Discussion

4 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?

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Discussion

5 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 - Human ileum using RNeasy Mini Kit from Qiagen.

Paper title

Lack of megalin expression in adult human terminal ileum suggests megalin‐independent cubilin/amnionless activity during vitamin B absorption

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Manufacturer protocol

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

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Videos

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