|
| Status |
Public on Dec 22, 2022 |
| Title |
DECB-6 |
| Sample type |
RNA |
| |
|
| Source name |
liver organization,35d,0.08mL/20g/d,replicate 1
|
| Organism |
Mus musculus |
| Characteristics |
tissue: liver
gender: male
|
| Treatment protocol |
After they were acclimated to the laboratory environment for 35d, 24 male mice were randomly divided into 2 groups: blank control group (CON), DECB group, 12 mice in each group. Mice in the CON group were given normal saline (0.08mL/20g), those in the DECB group were given DECB (0.08mL/20g) intragastrically once daily continuously for 35d.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total liver RNA was extracted with Trizol reagent (Invitrogen, Gaithersburg, MD, USA) and the mRNA was purified according to the instructions of RNeasy mini kit (Qiagen, Valencia, CA, USA). RNA samples were assessed for RNA integrity, inhibitor and DNA contamination by referring to the kit instructions. The quality of mRNA was detected by agarose gel electrophoresis, and the content was determined by UV spectrophotometer.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 700μL RNA using the RNeasy Mini Kit (Qiagen p/n 74104) according to the manufacturer's instructions, followed by RNAeasy column purification. The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000.
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| |
|
| Hybridization protocol |
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 at 60°C for 30 min, and 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.
|
| Scan protocol |
The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).
|
| Description |
Gene expression after 30d in mouse liver
|
| 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, genes that at least 3 out of 9 samples have flags in Detected (“All Targets Value”) were chosen for further data analysis. Differentially expressed genes with statistical significance between the two groups were identified through Volcano Plot filtering. Differentially expressed genes between the two samples were identified through Fold Change filtering. Hierarchical Clustering was performed using the R scripts. GO analysis and Pathway analysis were performed in the standard enrichment computation method.
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| |
|
| Submission date |
Dec 23, 2019 |
| Last update date |
Dec 22, 2022 |
| Contact name |
guangyu xu |
| E-mail(s) |
xuguangyu2005@163.com
|
| Organization name |
College of Pharmacy, Beihua University
|
| Street address |
Binjiang Road 3999,jilin city,132013
|
| City |
jilin city |
| ZIP/Postal code |
+860432 |
| Country |
China |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE142545 |
Effects of deproteinized extract of calf blood on the energy metabolism of acute hypoxia model mice and its targets |
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