Segments of jejunum were treated with RNAlater™ Stabilization Solution (Thermo-Fisher Scientific, Waltham, MA) for 24 hours at 4° C and stored at −80° C. Frozen segments of jejunum were thawed and lysed using QIAshredder homogenizer spin columns (QIAGEN, Venlo, Netherlands). Total RNA was extracted using the QIAGEN RNeasy mini kit (QIAGEN) according to the manufacturer’s instructions. Complementary DNA was synthesized from 0.5ug of total RNA using the iScript cDNA Synthesis kit (Bio-rad, Hercules, CA) following the manufacturer’s instruction. Messenger RNA quantification was measured using TaqMan Gene Expression Assays (Thermo-Fisher Scientific, Waltham, MA) using comparative Ct method and calculated in the open-source software package R (26 ).
Qiashredder homogenizer spin columns
Manufactured by Qiagen
Sourced in Canada
The QIAshredder homogenizer spin columns are a laboratory tool used for sample preparation. They are designed to efficiently homogenize and disrupt cells or tissue samples, releasing the cellular contents for subsequent processing or analysis.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy mini kit, by Qiagen (3 mentions)
Taqman gene expression assay, by Thermo Fisher Scientific (2 mentions)
Rnalater stabilization solution, by Thermo Fisher Scientific (1 mentions)
Iscript cdna synthesis kit, by Bio-Rad (1 mentions)
Omni tissue homogenizer, by Omni International (1 mentions)
Rlt lysis buffer, by Qiagen (1 mentions)
Hiseq 4000, by Illumina (1 mentions)
Truseq stranded mrna sample preparation kit, by Illumina (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Qiazol, by Qiagen (1 mentions)
Microarray suite 5, by Thermo Fisher Scientific (1 mentions)
Human genome u133 plus2.0 gene chip array, by Thermo Fisher Scientific (1 mentions)
3 protocols using qiashredder homogenizer spin columns
1
Quantifying Intestinal mRNA Levels
2
RNA Extraction and Sequencing from Organs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Organs (OB, BRN, CBL, BST, STM, LIV, and iBAT) were homogenized in Lysis RLT Buffer (Qiagen) supplemented with 1% β-mercaptoethanol (Sigma–Aldrich) by using the OMNI tissue homogenizer (OMNI International). Homogenized organ lysates were then loaded onto the QIAshredder homogenizer spin columns (Qiagen) for further homogenization and elimination of insoluble debris. Total RNA was extracted by using the RNeasy Mini Kit (Qiagen), according to the manufacturer's protocol. White adipose tissues (pgWAT and psWAT) were homogenized in Qiazol (Qiagen) and RNA extracted with the Lipid RNeasy Lipid Tissue Mini Kit (Qiagen). mRNA was prepared for sequencing by using the TruSeq stranded mRNA sample preparation kit (Illumina), with a selected insert size of 120–210 bp. All samples were sequenced on an Illumina HiSeq 4000, generating paired-end 150 bp sequencing reads, and had an average depth of 39.98 ± 1.05 (SEM) million reads (Additional file 2: Table S1 ).
3
RNA Isolation and Microarray Analysis
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