|
| Status |
Public on Jul 26, 2019 |
| Title |
2-1-4 |
| Sample type |
RNA |
| |
|
| Source name |
brain
|
| Organism |
Mus musculus |
| Characteristics |
tissue: hypothalamus
gender: Male
strain: Balb/c
age: 10 weeks
|
| Treatment protocol |
The mice were euthanized by cervical dislocation and brains were dissected using stereomicroscope to separate the four experimental brain regions (olfactory bulb, hypothalamus, hippocampus as well as cerebellum) according to mouse brain atlas
|
| Growth protocol |
10-week-old male BALB/c mice (Vital River Laboratories, Beijing, China) were used in this study and the mice were housed in a temperature-controlled room under a 12 hours light/12 hours dark cycle, with free access to food and water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the brain regions using TRIzol Reagent. RNA was purified from total RNA after removal of rRNA (mRNA-ONLY™ Eukaryotic mRNA Isolation Kit, Epicentre)
|
| Label |
Cy3
|
| Label protocol |
Each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3’ bias utilizing a random priming method (Arraystar Flash RNA Labeling Kit, Arraystar). 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.
|
| |
|
| Hybridization protocol |
1 μg of each labeled cRNA was fragmented by adding 5 μl 10 × Blocking Agent and 1 μl of 25 × Fragmentation Buffer, then heated the mixture at 60 °C for 30 min, finally 25 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven.
|
| Scan protocol |
The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).
|
| 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 v12.1 software package (Agilent Technologies). After quantile normalization of the raw data, LncRNAs and mRNAs have flags in Present or Marginal (“All Targets Value”) were chosen for further data analysis.
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| |
|
| Submission date |
Nov 13, 2017 |
| Last update date |
Jul 26, 2019 |
| Contact name |
xiaoyuan song |
| E-mail(s) |
songxy5@ustc.edu.cn
|
| Phone |
86-551-63607752
|
| Organization name |
University of Science & Technology of China
|
| Street address |
Huangshan Road
|
| City |
Hefei |
| ZIP/Postal code |
230027 |
| Country |
China |
| |
|
| Platform ID |
GPL19286 |
| Series (1) |
| GSE106832 |
Expression profile of lncRNAs and mRNAs in 4 brain regions of adult mouse |
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