RNeasy FFPE Kit

RNA isolation / purification Tissue - Human Nose

Experiment
RNA isolation / purification Tissue - Human Nose
Product
RNeasy FFPE Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
- To allow complete penetration by formalin, use tissue samples less than 5 mm thick
Protocol tips
- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C
Downstream tips
- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay

Publication protocol

When using paraffin embedded formalin tissue sections Qiagen RNeasy FFPE kit (Qiagen, Valencia, CA) was used according to the manufacturer's instructions to extract mRNA. For mRNA extraction for fresh tissues, specimens were placed into Tri-Reagent (Fisher Scientific) supplemented with glassbeads (1.0 mm diameter, BioSpec Products, Inc., Bartlesville, OK) immediately after collection and processed as recommended by the manufacturer. First-strand cDNA conversion was performed using iScript and real time PCR was conducted with SYBR Green according to manufacturers’ protocols (Bio-Rad Laboratories, Hercules, CA). The gene coding for the ribosomal protein RPLP0 was used as reference. Gene accession numbers were as follows: NM_001002.3 (RPLP0), NM_003101 (SOAT1), NM_004345 (LL37), NM_004942.2 (HBD2), and NM_0011081551.2 (HBD3). Primers had been designed by IDT (LL37 and SOAT1, Integrated DNA Technologies, Inc., San Diego, CA) or using NCBI Primer blast (RPLP0, HBD2, and HBD3). All PCR primers were purchased from IDT. Amplicons had been previously sequenced (DNA Core Facility, Beckman Research Institute, City of Hope, Duarte, CA, or Retrogen Inc., San Diego, CA) and were computer matched with the known cDNA sequences of each molecule.

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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

4 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 Nose using RNeasy FFPE Kit from Qiagen.

Paper title
Assessment of epithelial innate antimicrobial factors in sinus tissue from patients with and without chronic rhinosinusitis
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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy FFPE Kit below.

Download PDF Download manufacturer protocol

Videos

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