RNeasy Fibrous Tissue Mini Kit

RNA isolation / purification Tissue - Mouse Uterus

Experiment
RNA isolation / purification Tissue - Mouse Uterus
Product
RNeasy Fibrous Tissue Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
- This kit purifies RNA from up to 30 mg tissue
Downstream tips
- If there is low RNA yield,Repeat RNA elution, incubate the RNeasy Mini spin column on the benchtop for 10 min with RNase-free water before centrifuging.

- If there is low A260/A280 value use 10 mM Tris·Cl, pH 7.5, not RNase-free water, to dilute the sample before measuring purity.

- If RNA does not perform well in downstream experiments this could be due to salt carryover dyuring elution step, so ensure that Buffer RPE is at 15–25°C.

Publication protocol

Frozen uterine tissue ≤30 mg from CD-1 control and 30 ppm BPA exposure groups were homogenized for RNA isolation using an RNeasy Fibrous Tissue Mini Kit (Qiagen; Valencia, CA). Uterine RNA was converted to cDNA using a High Capacity cDNA Reverse Transcription Kit (Life Technologies; Grand Island, NY). Predesigned TaqMan RT-PCR assays (Applied Biosystems; Grand Island, NY) were used for the following genes: Col1a1 (Mm00801632_gH), Col3a1 (Mm00802296_g1), Mmp2 (Mm00439498_m1), Timp1 (Mm00441818_m1), Timp2(Mm00441825_m1), and Timp3(Mm00441826_m1). PCR amplification was performed in a final volume of 20 μL containing 1x TaqMan expression assay primers, Universal Master Mix (Applied Biosystems; Grand Island, NY), and 10 ng of cDNA. Amplification was performed in triplicates on a Step One Plus Real-Time PCR System (Applied Biosystems; Grand Island, NY) under the following fast conditions: 20 seconds at 95°C for one holding stage, then 1 second at 95°C and 20 seconds at 60°C for the cycling stage (60 cycles). Relative expression was quantified using the ΔΔCt method, using 18s as the endogenous control for normalization.

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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 - Mouse Uterus using RNeasy Fibrous Tissue Mini Kit from Qiagen.

Paper title
Strain-Specific Induction of Endometrial Periglandular Fibrosis in Mice Exposed During Adulthood to the Endocrine Disrupting Chemical Bisphenol A
Full paper
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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