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Status |
Public on Sep 23, 2021 |
Title |
2DRG24H |
Sample type |
RNA |
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Source name |
gene expression at 24h in cultured DRG neurons
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Organism |
Rattus norvegicus |
Characteristics |
strain: Sprague-Dawley gender: male tissue: Cultured DRG neurons timepoint: 24h gender: male
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Treatment protocol |
At 0h, 1.5h, 3h, 6h, 12h, 18h, 24h, 30h, 36h after neurons planted, the total RNA was isolated from these cells
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Growth protocol |
DRG neurons were obtained from the adult male Sprague-Dawley rats (180–220 g). After isolated from spinal cord, DRGs were rinsed twice in CMF-HBSS and then primary digesting with 0.1% collagenase type I and 0.25% trypsin. The single-cell suspension was spun for 5 min at 900 rpm with 15% BSA after triturated 3 times with 1 ml pipette tip. After discarding the supernatant, the cells were resuspended and planted with DMEM/F12 (Invitrogen) medium.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using TRIzol (Invitrogen) and miRNeasy mini kit (QIAGEN) according to manufacturer’s instructions, which efficiently recovered all RNA species, including miRNAs. RNA quality and quantity was measured by using nanodrop spectrophotometer (ND-1000, Nanodrop Technologies) and RNA Integrity was determined by gel electrophoresis.
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Label |
Cy3
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Label protocol |
Sample labeling and array hybridization were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology). Briefly, total RNA from each sample was linearly amplified and labeled with Cy3-UTP.
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Hybridization protocol |
The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000. 1 μg of each labeled cRNA was fragmented by adding 11 μl 10 × Blocking Agent and 2.2 μl of 25 × Fragmentation Buffer, then heated the mixture at 60 °C for 30 min, finally 55 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 100 μl of hybridization solution was dispensed into the gasket slide and assembled to the gene expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven. The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).
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Scan protocol |
Then the slides were scanned immediately with the Agilent DNA Microarray Scanner (part number G2505C).
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Data processing |
Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images. Quantile normalization and subsequent data processing were performed with using the GeneSpring GX v11.5.1 software package (Agilent Technologies) Quantile normalization was performed with using the GeneSpring GX v11.5.1 software package (Agilent Technologies)
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Submission date |
Sep 23, 2019 |
Last update date |
Sep 23, 2021 |
Contact name |
Lili Zhao |
E-mail(s) |
zhaoll@nicemice.cn
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Organization name |
Nantong university
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Street address |
19 Qixiu
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City |
Nantong |
ZIP/Postal code |
226000 |
Country |
China |
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Platform ID |
GPL14746 |
Series (2) |
GSE137838 |
Altered mRNA expression of cultured dorsal root ganglia neurons at different times of rat |
GSE137839 |
Altered miRNA and mRNA expression of cultured dorsal root ganglia neurons at different times of rat |
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