RNeasy Mini Kit

RNA isolation / purification Tissue - Mouse Spleen

Experiment
RNA isolation / purification Tissue - Mouse Spleen
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

Spleens were collected in RNA later (Ambion, Austin, TX, USA) and RNA isolated using RNeasy Mini kit following the manufacturer’s protocol (QIAGEN, Valencia, CA, USA). RNA (500 ng each) was reverse transcribed to cDNA using Superscript VILO cDNA Synthesis kit (Invitrogen Corporation, 11754–250, California, USA) and Real Time PCR was performed using SYBR Green Core Reagent kit and a 7300 RT-PCR system (Applied Biosystems, Austin, TX, USA). Primers are listed in S2 Table. GAPDH was used for normalization.

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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 Spleen using RNeasy Mini Kit from Qiagen.

Paper title
Serpin Treatment Suppresses Inflammatory Vascular Lesions in Temporal Artery Implants (TAI) from Patients with Giant Cell Arteritis
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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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