|
| Status |
Public on Jul 31, 2017 |
| Title |
B16F0 exosomes isolated from serum-free media media conditioned by B16F0 cells for 24 hr, rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
B16F0 exosomes
|
| Organism |
Mus musculus |
| Characteristics |
cell line: B16F0
sample type: exosomes isolated from B16F0 conditioned media
molecule subtype: exosomal RNA
|
| Growth protocol |
Cultured for 24 hours in serum-free media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA isolated from B16F0 exosomes and cells by RNeasy Plus kit (Qiagen) was quantified using Nanodrop and analyzed by on-chip-electrophoresis using the Agilent Bioanalyzer and was processed by the Ambion WT Expression Kit.
|
| Label |
biotin
|
| Label protocol |
Each reaction yielded between 6 to 9 micrograms of cDNA, whereby 5.5 mug of cDNA was fragmented and labeled with biotin by the GeneChip WT Terminal Labeling Kit (Affymetrix). The efficiency of fragmentation reaction was checked via Agilent Bioanalyzer.
|
| |
|
| Hybridization protocol |
The entire reaction of fragmented and biotin-labeled cDNA (50 mu) with added hybridization controls was hybridized to the mouse GeneChip 1.0 ST Exon Arrays (Affymetrix) at 45oC for 17 hours in GeneChip Hybridization Oven 640 (Affymetrix).
|
| Scan protocol |
Mouse GeneChip 1.0 ST Exon Arrays were stained using FS 450_0001 protocol in Affymetrix GeneChip Fluidics Station 450. Phycoerythrin labeling was detected within the Affymetrix GeneChip Scanner 3000 7G plus using 532 nm light and detected by a photomultiplier tube.
|
| Description |
6 exosome 24h_(MoEx-1_0-st-v1)
|
| Data processing |
Expression Console software (Affymetrix) was used to check quality controls of hybridized chips. All chips that passed quality controls were RMA normalized using Expression Console software.
probe group file: MoEx-1_0-st-v1.r2.pgf
meta-probeset file: MoEx-1_0-st-v1.r2.dt1.mm8.mps
|
| |
|
| Submission date |
Jul 31, 2015 |
| Last update date |
Jul 31, 2017 |
| Contact name |
David John Klinke |
| E-mail(s) |
David.Klinke@mail.wvu.edu
|
| Phone |
304-293-9346
|
| Organization name |
West Virginia University
|
| Department |
Chemical Engineering
|
| Street address |
P.O. Box 6102
|
| City |
Morgantown |
| State/province |
WV |
| ZIP/Postal code |
26506-6102 |
| Country |
USA |
| |
|
| Platform ID |
GPL6096 |
| Series (2) |
| GSE71610 |
Expression data from parental B16F0 cells and B16F0 exosomes [MoEx] |
| GSE102951 |
Expression data from parental B16F0 cells and B16F0 exosomes |
|
