TRIzol (Invitrogen) was used for total RNA extractions according to the manufacturer’s protocol. RNA quantity and quality were measured by NanoDrop ND-1000. Its OD260/OD280 ratio was confirmed to be higher than 1.8. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. The RNA samples were sent to Kangchen Biotechnology Limited Company (Shanghai, China) for hybridization to the Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA). Each RNA sample was hybridized to one microarray slide. 1 μg of total RNA from each sample was amplified and transcribed into fluorescent cRNA with using the manufacturer’s Agilent’s Quick Amp Labeling protocol (version 5.7, Agilent Technologies). The labeled cRNAs were hybridized onto the Whole Genome Oligo Array (4x44K, Agilent Technologies).
Agilent sheep gene expression microarray
Manufactured by Agilent Technologies
Sourced in United States
The Agilent sheep gene expression microarray is a laboratory instrument designed for analyzing gene expression patterns in sheep samples. It provides a comprehensive platform for studying the activity and regulation of genes in sheep across a wide range of biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Trizol, by Thermo Fisher Scientific (2 mentions)
Kreatech uls labeling kit, by Agilent Technologies (2 mentions)
Agilent c scanner 20 bit, by Agilent Technologies (2 mentions)
Rna clean and concentrator spin columns, by Zymo Research (2 mentions)
Agilent bioanalyzer, by Agilent Technologies (2 mentions)
Quick amp labeling protocol, by Agilent Technologies (1 mentions)
Whole genome oligo array, by Agilent Technologies (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
4 protocols using agilent sheep gene expression microarray
1
Sheep Transcriptome Analysis Using Microarray
2
Sheep Gene Expression Microarray Analysis
3
RNA Extraction and Microarray Profiling
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Total RNA was extracted using TRIzol (Invitrogen). The extraction was performed according to the manufacturer’s protocol28 (link). RNA integrity and concentration were evaluated by Nanodrop. Our experiments were accomplished at 2010, when Agilent 2100 Bioanalyzer were not available in China. So we deployed Nanodrop and agarose gel electrophoresis to detect the quality of RNA samples. The hybridization of the RNA samples to the Agilent Sheep Gene expression Microarray (Santa Clara, CA, USA)28 (link) was performed by the Kangchen Biotechnology Limited Company (Shanghai, China).
4
Sheep Gene Expression Microarray Analysis
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Frozen tissues were shipped to MOgene, Inc. (St. Louis, MO) on dry ice. High quality RNA was isolated (RNA Integrity Number of 7.5-10 and 28S/18S ratio of 1.6-2.1) and confirmed with an Agilent Bioanalyzer at MOgene, Inc. Five hundred ng of total RNA was amplified using Agilent QuickAmp Labeling Kit (no dye) and purified using Zymo Research RNA Clean and Concentrator spin columns. Three μg of amplified RNA was Cy-labeled using the Kreatech ULS Labeling Kit, fragmented according to Agilent specifications and hybridized to the Agilent sheep gene expression microarray (Agilent Technologies, Santa Clara CA) for 17 hours at 65°C and 10 rpm. Slides were scanned on an Agilent C scanner (20 bit) at 5 μm resolution. Sixteen microarray hybridizations were performed, each with samples from an ANG II, losartan infused fetus (n = 4 for each), or a paired twin control (n = 8), using a mixed design on 2 slides with 8 microarrays per slide. Microarray data were submitted to the NCBI Gene Expression Omnibus (GEO), and can be accessed at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE45463 (project accession no. GSE45463).
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