|
| Status |
Public on Aug 04, 2006 |
| Title |
Transit amplifying cell sample 1 (B) |
| Sample type |
RNA |
| |
|
| Source name |
Cultured human keratinocytes at passage 3
|
| Organism |
Homo sapiens |
| Characteristics |
Human epidermal keratinocyte from healthy donor
|
| Treatment protocol |
Keratinocyte were picked upon putting them in suspension and seeded in single cell amplification buffer
|
| Growth protocol |
Keratinocytes were cultured on a feeder layer of J2-3T3 cells in FAD medium (1 part HAM’S F12, 3 parts Dulbecco’s modified Eagle’s medium, 1.8x10-4 M adenine), supplemented with 10% fetal calf serum (FCS), 0.5 µg/ml hydrocortisone, 5 µg/ml insulin, 10-10 M cholera enterotoxin and 10ng/ml EGF
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Single cells were seeded in amplification buffer
|
| Label |
biotin
|
| Label protocol |
amplified mRNA from single cultured epidermal keratinocytes
Biotinylated cDNA were prepared according to Tietjen et al. 2003, Neuron.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 microg of cDNA were hybridized for 16 hr at 45C on GeneChip Human Genome Array HU133. GeneChips were washed and stained using an Affymetrix GeneChip Instrument System.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner.
|
| Description |
Gene expression data from single cultured human epidermal keratinocytes
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings.
|
| |
|
| Submission date |
May 17, 2006 |
| Last update date |
Aug 04, 2006 |
| Contact name |
Kim B. Jensen |
| E-mail(s) |
[email protected]
|
| Organization name |
Cancer Research UK
|
| Department |
London Research Institute
|
| Lab |
Keratinocyte
|
| Street address |
44 Lincoln's Inn Fields
|
| City |
London |
| ZIP/Postal code |
WC2A 3PX |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL97 |
| Series (1) |
| GSE4858 |
Single cell expression profiling of human epidermal stem and transit amplifying cells |
|
