|
| Status |
Public on Apr 03, 2022 |
| Title |
CybbKO RPM F4/80high 3 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse resident peritoneal macrophage (B220-CD115+ F4/80high MHCII-)
|
| Organism |
Mus musculus |
| Characteristics |
genotype: CGD
tissue: Peritoneal lavage
gender: Male
age: 9wks
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNAeasy micro plus
|
| Label |
Cy5
|
| Label protocol |
RNA samples were amplified using SIGMA WTA2 RNA amplification system. Amplified cDNAs were chemically labeled with Kreatech ULS RNA labeling kit (Kreatech Diagnostics). Per reaction, 3ug of each RNA (+water=16ul) was mixed with Kreatech 10x labeling buffer (2ul) and Kreatech cy5/DY-ULS (2ul). The reactions were incubated at 85C for 15 minutes in the dark and placed on ice for 3 minutes. Labeled cDNA was purified with Qiagen PCR purification columns according to manufacturer’s protocol. cDNAs were quantitated on a Nanodrop spectrophotometer. Detailed protocol can be found at http://www.kreatech.com/fileadmin/user_upload/Documenten/PDF/12_man_EA-021-022-023__D0.6.pdf
|
| |
|
| Hybridization protocol |
The balanced aRNAs were suspended in Agilent 2X Gene Expression buffer (55ul), Agilent 10X Blocking agent (11ul), and Kreablock (6ul). The hybridization solutions were applied to Agilent Mouse v2 4x44K microarrays (G4846A-026655). Hybridization was carried out at 65C for 20 hours. Washing procedures were carried out according to Agilent gene expression protocols.
|
| Scan protocol |
Slides were scanned on an Agilent C-class Microarray scanner to detect Cy5 fluorescence, according to manufacturer's specifications.
|
| Description |
No treatment
|
| Data processing |
Gridding and analysis of images was performed using Feature Extraction (v11.5.1.1, Agilent Technologies). The extracted signal was quantile-normalizated using Partek.
|
| |
|
| Submission date |
Mar 16, 2022 |
| Last update date |
Apr 03, 2022 |
| Contact name |
Mary C Dinauer |
| E-mail(s) |
[email protected]
|
| Organization name |
Washington University In St Louis
|
| Department |
Pathology & Immunology
|
| Street address |
660 South Euclid Ave
|
| City |
St Louis |
| State/province |
Missouri |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL11202 |
| Series (2) |
| GSE198771 |
Analysis of resident peritoneal macrophage's transcriptome in WT and CGD mice |
| GSE198778 |
The leukocyte NADPH oxidase 2 (NOX2) plays a key role in pathogen killing and immunoregulation |
|
