QIAzol Lysis Reagent

RNA isolation / purification Tissue - Rat Lung

Experiment
RNA isolation / purification Tissue - Rat Lung
Product
QIAzol Lysis Reagent from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
- To promote dissociation of nucleoprotein complexes, place the sample at room temperature (15–25°C) for 5 min after homogenization
Downstream tips
- Recommends for a cleanup of the redissolved RNA using RNeasy® Kits, which are based on silica-membrane technology, in order to remove any contaminating phenol

- If A260/A280 is low or if DNA containation is high, reduce the amount of starting material and/or increase the volume of QIAzol Lysis Reagent and the homogenization time

Publication protocol

In a first step, four rat PCLS per tube were used for RNA isolation. Several commercially available kits were used for this according to the suppliers’ instructions: RNeasy Mini Kit (Qiagen, Hilden, Germany), QIAzol® (Qiagen, Hilden, Germany), MagJET (Thermo Fisher Scientific, Dreieich, Germany), and MagMAX™ (Ambion™/Thermo Fisher Scientific, Dreieich, Germany). Disruption and homogenization of PCLS in the respective solutions was performed using an Ultra-Turrax® (T8, IKA, Stauffen, Germany).

In a second step, optimized RNA extraction was achieved as follows: two PCLS were pooled, followed by disruption and homogenization of PCLS in 400 µL RLT lysis buffer (Qiagen, Hilden, Germany) using an Ultra-Turrax®. The homogenate was transferred to 1 volume of phenol/chloroform, carefully shaken for 30 s, and centrifuged for 5 min at 12,000×g. Subsequently, 1 volume of chloroform/isoamyl alcohol was added, again carefully shaken for 30 s, and centrifuged for 5 min at 12,000×g. The aqueous phase was transferred and RNA was cleaned up with MagMAX™ magnetic beads including the spin procedure step according to the supplier’s instructions. Total RNA was dissolved in RNase-free water and stored at −80 °C. For some samples an additional clean up step was performed using the RNeasy Mini Kit clean up protocol (Qiagen, Hilden, Germany).

RNA from A459 cells was isolated with the commercially available RNeasy Mini Kit according to the supplier’s instructions.

Full paper   Login or join for free to view the full paper.

Reviews

QIAzol Lysis Reagent from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

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?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Rat Lung using QIAzol Lysis Reagent from Qiagen.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Qiagen for QIAzol Lysis Reagent below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Tissue - Rat Lung using QIAzol Lysis Reagent from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms