RNeasy Mini Kit

RNA isolation / purification Tissue - Mouse Nose

Experiment

RNA isolation / purification Tissue - Mouse Nose

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

At day 21 in ALI culture, primary NECs were collected from the Transwell inserts with trypsin/EDTA and homogenized in lysis buffer from the Qiagen Mini RNeasy Kit (Qiagen, Germantown, Md). Nasal mucosal biopsy specimens were snap-frozen immediately after isolation. First-strand cDNA was reverse transcribed with SuperScript II (Invitrogen, Carlsbad, Calif) with 1 μg of total RNA, according to the manufacturer's instructions. RT-qPCR was performed for claudin-1, claudin-4, occludin, and ZO-1 and the housekeeping genes encoding β-actin (ACT2B) and β2-microglobulin. cDNA plasmid standards, consisting of purified plasmid DNA specific for each target, were used to quantify the target gene in the unknown samples

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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 - Mouse Nose using RNeasy Mini Kit from Qiagen.

Paper title

Impaired barrier function in patients with house dust mite-induced allergic rhinitis is accompanied by decreased occludin and zonula occludens-1 expression.

Full paper

Manufacturer protocol

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

Download manufacturer protocol

Videos

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