RNA was extracted from micropunches using TRIzol LS Reagent (Qiagen) followed by a Qiagen MagAttract RNA Universal Tissue M48 Kit customized protocol on the upper aqueous phase. First, 5ul of MagAttract Suspension E silica beads were added and mixed into each sample. Suspension E beads were separated from the mixture using a magnetic tube rack and 100uL of Wash Buffer MW was added. This process was repeated with another wash of Wash Buffer MW. Suspension E beads were again separated from the mixture and the wash buffer was aspirated. 100uL of Wash Buffer RPE was added, mixed, and aspirated twice. Isolated RNA was eluted from Suspension E beads in 25uL of RNase free water.
Magattract rna universal tissue m48 kit
Manufactured by Qiagen
Sourced in Germany
The MagAttract RNA Universal Tissue M48 Kit is a laboratory equipment product designed for the isolation of total RNA from various tissue samples. The kit utilizes magnetic bead-based technology to capture and purify RNA, providing a convenient and efficient method for RNA extraction.
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Lab products found in correlation
Trizol ls reagent, by Qiagen (1 mentions)
2100 bioanalyzer, by Agilent Technologies (1 mentions)
Nanodrop, by Thermo Fisher Scientific (1 mentions)
Beadarray reader, by Illumina (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Ht12 beadchip gene expression array, by Illumina (1 mentions)
2 protocols using magattract rna universal tissue m48 kit
1
RNA Extraction from Tissue Micropunches
2
RNA Extraction and Expression Analysis
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RNA was extracted from 28 tumors using MagAttract RNA Universal Tissue M48 Kit (Qiagen, Hilden, Germany) according to manufacturer’s protocol using BioRobot M48. RNA concentrations were determined by spectrophotometry (NanoDrop, Thermo Scientific, Wilmingron, DE, USA) and quality was analysed using the 2100 Bioanalyzer (Agilent technologies, Santa Clara, CA, USA).
Two hundred ng of total RNA from each sample was used for cRNA production by the Illumina TotalPrep RNA amplification kit (Ambion Inc., St. Austin, TX, USA) according to the provided protocol. The quality of purified cRNA was evaluated using the RNA 6000 p kit (Agilent Technologies) in the Agilent 2100 Bioanalyzer (Agilent Technologies). A total of 750 ng biotinylated cRNA was hybridized to the human HT12 Illumina Beadchip gene expression array (Illumina, San Diego, CA, USA) according to manufacturer’s protocol and scanned using the Illumina Bead Array Reader (Illumina). Expression array data was analysed using the Illumina BeadStudio V2011.1 software and samples were normalized using the cubic spline algorithm. Gene expression levels of the MX2 (ILMN_2231928), SMAD6 (ILMN_1767068) and SOCS3 (ILMN_1781001) genes were extracted from the arrays.
Two hundred ng of total RNA from each sample was used for cRNA production by the Illumina TotalPrep RNA amplification kit (Ambion Inc., St. Austin, TX, USA) according to the provided protocol. The quality of purified cRNA was evaluated using the RNA 6000 p kit (Agilent Technologies) in the Agilent 2100 Bioanalyzer (Agilent Technologies). A total of 750 ng biotinylated cRNA was hybridized to the human HT12 Illumina Beadchip gene expression array (Illumina, San Diego, CA, USA) according to manufacturer’s protocol and scanned using the Illumina Bead Array Reader (Illumina). Expression array data was analysed using the Illumina BeadStudio V2011.1 software and samples were normalized using the cubic spline algorithm. Gene expression levels of the MX2 (ILMN_2231928), SMAD6 (ILMN_1767068) and SOCS3 (ILMN_1781001) genes were extracted from the arrays.
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