RNeasy Mini Kit

RNA isolation / purification Tissue - Human Brain

Experiment
RNA isolation / purification Tissue - Human Brain
Product
RNeasy Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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

Total RNA was extracted from frozen PT biopsy samples using RNeasy minikits and the RNAse-Free DNAse Set (Qiagen, Germantown, MA). The RNA quantity and quality were determined with Nanodrop and Agilent analyzers. RNA from postmortem human brain tissue was used as positive control. Eight hundred micrograms of RNA were used for cDNA synthesis using the AMV First-Strand cDNA Syhthesis Kit (no. 12328-040; Invitrogen, Carlsbad, CA). Two microliters of cDNA synthesis reaction were used in a 25-μl real-time PCR reaction with FAM-labeled Taqman Gene Expression Assays for NR-2A (NMDAR2a; Hs00168219_m1) and MAP2 (Hs01103234_g1; Applied Biosystems, Foster City, CA). β-Actin (Hs03023943_g1) was used as the reference control gene. The amplification conditions were: 10 min at 95°C followed by 40 cycles of 15 s at 95°C, 1 min at 60°C. CT values were analyzed using the 2-ΔΔCT method.

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Discussion

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?

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 Brain using RNeasy Mini Kit from Qiagen.

Paper title
Brain-derived antigens in lymphoid tissue of patients with acute stroke.
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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