RNeasy Mini Kit

RNA isolation / purification Tissue - Human Iris

Experiment

RNA isolation / purification Tissue - Human Iris

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

Iris specimens were lysed by sonication and total RNA recovered with Trizol Reagent (Invitrogen Corp, CA, USA) and the RNeasy kit (Qiagen, Valencia, CA) as described previously.17 Five nanograms of total RNA was reverse‐transcribed into cDNA using random hexamer primers (Invitrogen Corp) with Superscript III reverse transcriptase (Invitrogen Corp). All qPCR reactions, comprising the Power SYBR Green PCR Master Mix (Applied BioSystems, CA, USA), were performed in triplicates in volumes of 10 µl in 384‐well microtiter plates and run using the Roche LightCycler 480 System (Roche Diagnostics Corp, Indianapolis, USA). All mRNA levels were measured as CT threshold levels. ACTB was used as the housekeeping gene, determined to be the most suitable of four analysed (ACTB, RNA18S1, GAPDH, RPL13A) using the NormFinder software

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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 Iris using RNeasy Mini Kit from Qiagen.

Paper title

Distinct iris gene expression profiles of primary angle closure glaucoma and primary open angle glaucoma and their interaction with ocular biometric parameters

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Manufacturer protocol

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

Download manufacturer protocol

Videos

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