|
| Status |
Public on Aug 15, 2016 |
| Title |
Day 4 Control |
| Sample type |
RNA |
| |
|
| Source name |
Primary murine cells
|
| Organism |
Mus musculus |
| Characteristics |
strain: Balb/c
cell type: Macrophage-Osteoclast
|
| Treatment protocol |
Mtb infections were carried out by infecting primary cells with Mtb at MOI of 1:5 for 4hours.
|
| Growth protocol |
Murine bone marrow derived macrophage precursors were initially grown in presence of MCSF while post Mtb treatment cells were cultured with MCSF and RANKL growth factors.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from uninfected and Mtb infected cells was isolated by TRIzol RNA isolation method and later purified by RNAeasy column purification.
|
| Label |
Biotin
|
| Label protocol |
300ng RNA was taken as input RNA and was processed with Ambion WT Expression Kit.cDNAS were fragmented and labeled with Affymetrix terminal labeling kit.
|
| |
|
| Hybridization protocol |
Labeled fragments were hybridized to Affymetrix WT Mouse ST2.0 gene array for 16hr at 45°C with 60rpm in Affymetrix Hybridization oven 645.
|
| Scan protocol |
Hybridized microarrays were washed and stained in Affymetrix fluidic station 450 and scanned in Affymetrix G3000 Gene Array Scanner.
|
| Description |
Poly-A controls were added to input RNA.
|
| Data processing |
Image analysis of microarray chips was carried out by Affymetrix Genechip command console software which uses MAS5 algorithm. Affymetrix Expression Console was used to analyze ,CEL files to generate normalized signal intensities form raw signal intensities using RMA algorithm.
|
| |
|
| Submission date |
Sep 18, 2014 |
| Last update date |
Aug 15, 2016 |
| Contact name |
Kuldeep Singh Attri |
| E-mail(s) |
kuldeep.attri@igib.in
|
| Phone |
+91-9953322057
|
| Organization name |
CSIR-IGIB
|
| Department |
Systems Biology
|
| Lab |
Rajesh Gokhale Lab
|
| Street address |
Sukhdev Vihar
|
| City |
New Delhi |
| State/province |
Delhi |
| ZIP/Postal code |
110020 |
| Country |
India |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE61531 |
Gene expression based profiling of Mycobacterium tuberculosis infected primary murine osteoclasts. |
|
