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| Status |
Public on Jan 02, 2024 |
| Title |
Fscn2-/- mouse_8 weeks of age 3 |
| Sample type |
RNA |
| |
|
| Source name |
inner ears, NO. 6 mouse (No gender limitation)
|
| Organism |
Mus musculus |
| Characteristics |
tissue: inner ear
genotype: Fscn2-/-
|
| Treatment protocol |
The Fscn2 knockout mice were generated on the C57BL/6J background from Cyagen Biosciences (Guangzhou, China). The Fscn2 null mutation was made using the TALEN technique, with two vectors targeting the first exon of Fscn2 The wild-type mice (Fscn2+/+), heterozygous mice (Fscn2+/-), and homozygous mice (Fscn2-/-) were genotyped using a polymerase chain reaction (PCR).
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| Growth protocol |
Experimental mice were bred in a specific pathogen-free animal facility at Binzhou Medical University. The animal studies were conducted in accordance with the principles set forth in the Guide for the Care and Use of Laboratory Animals of Binzhou Medical University and were approved by that university’s Institutional Animal Use and Care Committee (protocol 14-0514)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was quantified by the NanoDrop ND-2000 (Thermo Scientific)and the RNA integrity was assessed using Agilent Bioanalyzer 2100 (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 μg RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer
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| |
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| Hybridization protocol |
0.6 μg of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/μg cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 22.5μl containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers' instructions. On completion of the fragmentation reaction, 22.5μl of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Mouse lncRNA V3 (4*180K) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation
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| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x180k array slides (Scan Area 61x21.6 mm, Scan resolution 3um, Dye channel is set to Green and Green PMT is set to 100%)
|
| Description |
Gene expression of inner ears from Fscn2-/- mouse_8 weeks of age 3
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities as the raw data. Raw data were normalized in quantile algorithm with Genespring 13.0(Agilent). Probe that at least 1 out of 2 samples flagged as Detected were maintained.
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| |
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| Submission date |
Dec 28, 2023 |
| Last update date |
Jan 02, 2024 |
| Contact name |
Wenjing Shang |
| E-mail(s) |
wenjshang@126.com
|
| Organization name |
Binzhou Medical University
|
| Street address |
NO.346 Guanhai Road
|
| City |
Yantai |
| State/province |
Shandong |
| ZIP/Postal code |
364000 |
| Country |
China |
| |
|
| Platform ID |
GPL25663 |
| Series (1) |
| GSE252226 |
Gene expression profiles in the inner ears of Fscn2 knockout mice compared with the control mice |
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