|
| Status |
Public on Feb 11, 2019 |
| Title |
per2 from RAW264.7 cells infected with B. melitensis M5-90 ∆per for 4h |
| Sample type |
RNA |
| |
|
| Source name |
RAW264.7 cells
|
| Organism |
Mus musculus |
| Characteristics |
cell line: RAW264.7 cells
infection condition: B. melitensis M5-90 delta-per
infection time: 4h
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the Animal Tissue RNA Purification Kit (Norgen Biotek Corporation, Thorold, Canada), following the manufacturer's recommended protocol.
|
| Label |
cy3
|
| Label protocol |
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.
|
| |
|
| 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 chip 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.
|
| 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 |
153 differentially expressed genes were detected in this microarray
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software (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(Agilent).
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| |
|
| Submission date |
Feb 10, 2019 |
| Last update date |
Feb 11, 2019 |
| Contact name |
Xie qin |
| E-mail(s) |
[email protected]
|
| Phone |
15618926050
|
| Organization name |
LC sciences
|
| Street address |
Minchi first road, pujiang town, minhang district, Shanghai
|
| City |
shanghai |
| ZIP/Postal code |
200000 |
| Country |
China |
| |
|
| Platform ID |
GPL11202 |
| Series (2) |
| GSE126343 |
The role of mirna-146b-tbc1d14 in the autophagy of RAW264.7 cells mediated by B. melitensis m5-90 [mRNA] |
| GSE126499 |
The role of mirna-146b-tbc1d14 in the autophagy of RAW264.7 cells mediated by B. melitensis m5-90 |
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