RNeasy Mini Kit

RNA isolation / purification Tissue - Mouse Vagina

Experiment
RNA isolation / purification Tissue - Mouse Vagina
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

Following collection, RNA in vaginal wash fluid was stabilized by addition of 10 volumes of RNAlater solution (Qiagen, Valencia, CA), and samples were immediately processed or frozen at −80°C for later use. Total RNA was isolated from vaginal wash fluid or homogenized vaginal tissue by shaking with 0.1-mm silica spheres using Lysing Matrix B FastRNA tubes and a FastPrep high-speed reciprocating shaking device (QBioGene, Carlsbad, CA). RNA was further purified by use of an RNeasy minikit (Qiagen). Genomic DNA was removed by digestion during isolation (RNase-free DNase set; Qiagen) and by treatment with DNase I (amplification grade; Invitrogen) following spin-column purification. The integrity of the RNA was assessed by electrophoresis, and purity was measured by determination of the A260/A280 ratio (purity was acceptable if it was >1.8).

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Discussion

Discussion

2 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

3 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 - Mouse Vagina using RNeasy Mini Kit from Qiagen.

Paper title
Murine Vaginal Colonization Model for Investigating Asymptomatic Mucosal Carriage of
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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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