RNeasy Mini Kit

RNA isolation / purification Tissue - Rat Bone

Experiment
RNA isolation / purification Tissue - Rat Bone
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

A combination of TRIzol method and RNeasy Mini Kit (QIAGEN, Sollentuna, Sweden) were used to isolate RNA from bone samples (Reno et al., 1997). Frozen bone samples were pulverized by tungsten balls, inside vials cooled by liquid Nitrogen, in a Retsch mixer mill MM 200 (Retsch, Haan, Germany). Samples were kept frozen during the whole homogenization procedure. Immediately after pulverization, TRIzol (Invitrogen) was added to samples and left to thaw at room temperature. Next, chloroform wad added followed by centrifugation and the aqueous phase was then transferred to new tubes and mixed gently with 70% ethanol. RNA samples were purified according to the RNeasy Mini Kit instructions. RNase-free DNase Set (QIAGEN, Sollentuna, Sweden) was used to exclude possible DNA contamination. Quality of RNA samples including concentration and RIN (RNA Integrity Number) value were checked by Nanodrop ND-1000 (NanoDrop Technologies, Wilmington, DE) and Agilent RNA 6000 Nano Kit (Agilent Technologies, Böblingen, Germany). Samples were kept at − 80 °C before microarray analysis.

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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 - Rat Bone using RNeasy Mini Kit from Qiagen.

Paper title
Mechanical instability and titanium particles induce similar transcriptomic changes in a rat model for periprosthetic osteolysis and aseptic loosening
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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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