Approximately 10% of each lobe of lung was snap-frozen at -80°C to preserve RNAs. The lungs were then homogenized in Trizol solution (Qiagen, Hilden, Germany) using a gentleMACS tissue dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany), and total RNA was immediately extracted using the miRNeasy miRNA extraction kit (Qiagen). The isolated total RNA samples containing miRNAs were then subjected to miRNA and gene microarray analyses.
Mirneasy mirna extraction kit
Manufactured by Qiagen
Sourced in Germany
The MiRNeasy miRNA extraction kit is a product offered by Qiagen that is designed for the isolation and purification of microRNA (miRNA) from a variety of sample types, including cells, tissues, and body fluids. The kit utilizes Qiagen's proprietary technology to efficiently capture and extract miRNA molecules from the sample, providing a reliable and consistent method for miRNA analysis.
Automatically generated - may contain errors
3 protocols using mirneasy mirna extraction kit
1
Lung tissue RNA extraction and analysis
2
Plasma RNA Extraction for miRNA Analysis
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Plasma samples were thawed on ice and then further centrifuged 3,000g for 5 min at 4°C in order to completely remove cell debris. MiRNeasy miRNA extraction kit (Qiagen, Hilden, Germany) was used to extract RNA from 50 µl of plasma from each patient, by adding 1,200 µl of Trizol LS (Lifetechnologies, CA, USA) and finally eluted in 30 µl of nuclease-free water. Moreover, 25 pmol of the spike-in control ath-miR-159a were added to each sample, in order to control variations in RNA extraction procedures.
3
Serum RNA Extraction for Biomarker Analysis
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Serum samples were thawed on ice and then further centrifuged 3000× g for 5 min at 4 °C in order to completely remove contaminant cells and cell debris. A total of 50 μL of serum from each patient was diluted in 350 μL of nuclease-free water to avoid protein aggregates. Then, RNA was extracted from diluted serum by using miRNeasy miRNA extraction kit (Qiagen, Hilden, Germany), by adding 1.2 mL of Trizol LS (Lifetechnologies, Carlsbad, CA, USA) as lysis buffer and finally eluted in 30 μL of nuclease-free water.
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