RNeasy Mini Kit

RNA isolation / purification Tissue - Human Breast

Experiment

RNA isolation / purification Tissue - Human Breast

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

The central section of fresh-frozen tissues was homogenized using a MagNA Lyser homogenizer (Roche), and RNA was isolated by QIAzol extraction followed by purification on an RNeasy column as described previously [5]. RNA quality and quantity were assessed on an Agilent 2100 Bioanalyzer and a ND-1000 NanoDrop spectrophotometer, respectively. Two-color 4 × 44 K Agilent whole-genome arrays were run on the RNA samples. Data were preprocessed before analytic statistical analyses: lowess-normalization, setting value of the probes that had a signal less than 10 dpi in either channel as missing, excluding probes that had more than 20% missing data across all samples, imputing the missing values using k-nearest neighbors' imputation (with k = 10), and collapsing the duplicate probes by averaging. Microarray data used in these analyses will be publicly available through the Gene Expression Omnibus (accession number GSE111601) www.ncbi.nlm.nih.gov/geo/.

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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 Breast using RNeasy Mini Kit from Qiagen.

Paper title

Correlated metabolomic, genomic, and histologic phenotypes in histologically normal breast tissue

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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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