RNeasy Mini Kit

RNA isolation / purification Tissue - Human Hippocampus

Experiment
RNA isolation / purification Tissue - Human Hippocampus
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

Brain samples were homogenized in Trizol and total RNA was isolated from cells using the RNeasy Kit (Qiagen) according to the manufacturer's protocol. An additional DNase1 digestion step was performed to ensure that the samples were not contaminated with genomic DNA. RNA purity was checked by calculating RIN in Agilent BioAnalyzer 2100 (Agilent, Santa Clara, CA). Transcriptome sequencing libraries were prepared using the TruSeq RNA Access Library Prep Kit® (Illumina, CA; US). Paired end sequencing (100 bp) was performed at an Illumina HiSeq 2500 instrument. All the 100 bp paired end raw reads were quality checked for low quality bases and adapter sequences (Processing of raw reads). Quality check was done using NGS QC Toolkit v2.3.1 [58]. Low quality bases and reads were filtered out from the datasets prior to read mapping. We used Tophat RNA aligner to map sequence read to UCSC (the University of California Santa) reference human genome Hg19

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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

4 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 Hippocampus using RNeasy Mini Kit from Qiagen.

Paper title
RNA-seq analysis of hippocampal tissues reveals novel candidate genes for drug refractory epilepsy in patients with MTLE-HS.
Full paper
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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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