|
Status |
Public on May 01, 2016 |
Title |
Fibroblast cell line F3, cell 6, sister cell 1 |
Sample type |
genomic |
|
|
Channel 1 |
Source name |
Reference DNA
|
Organism |
Homo sapiens |
Characteristics |
sample type: reference gender: Male
|
Treatment protocol |
Single fibroblasts were plated separately and the two cells that derived following a cell division were isolated individually.
|
Growth protocol |
Primary fibroblasts were grown in 75-cm2 plastic flasks (BD Falcon, USA) under standard culture conditions in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12) (Gibco, USA) medium complemented with 10% Fetal Bovine Serum (FBS) (Thermo Scientific, USA).
|
Extracted molecule |
genomic DNA |
Extraction protocol |
The genome of collected single cells was amplified using the Sureplex amplification system (BlueGnome/Illumina, USA).
|
Label |
Cy3
|
Label protocol |
The fluorescent labeling system (BlueGnome/Illumina) was used for sample labeling. Samples and Reference DNAs were labeled using Cy5 or Cy3 for 2-4h at 37░C. Samples were combined with the respective references prior to hybridization.
|
|
|
Channel 2 |
Source name |
Primary single fibroblast, F3
|
Organism |
Homo sapiens |
Characteristics |
cell type: primary single fibroblast gender: Female
|
Treatment protocol |
Single fibroblasts were plated separately and the two cells that derived following a cell division were isolated individually.
|
Growth protocol |
Primary fibroblasts were grown in 75-cm2 plastic flasks (BD Falcon, USA) under standard culture conditions in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12) (Gibco, USA) medium complemented with 10% Fetal Bovine Serum (FBS) (Thermo Scientific, USA).
|
Extracted molecule |
genomic DNA |
Extraction protocol |
The genome of collected single cells was amplified using the Sureplex amplification system (BlueGnome/Illumina, USA).
|
Label |
Cy5
|
Label protocol |
The fluorescent labeling system (BlueGnome/Illumina) was used for sample labeling. Samples and Reference DNAs were labeled using Cy5 or Cy3 for 2-4h at 37░C. Samples were combined with the respective references prior to hybridization.
|
|
|
|
Hybridization protocol |
200 ng of sample DNA and equal amount of commercially available reference DNA of the were labeled for 2 h by random primer labeling (BlueGnome/Illumina, USA) using Cy5- and Cy3-dCTPs, respectively.
|
Scan protocol |
DNA-microarray scanner (Agilent Technologies, UK).
|
Description |
Sureplex whole genome amplified single-cell DNA 131
|
Data processing |
Separation of the duplex raw images by ImageViewer and the analysis of the data by BlueFuse v3.0 software (BlueGnome/Illumina, USA).
|
|
|
Submission date |
Nov 16, 2015 |
Last update date |
May 01, 2016 |
Contact name |
Eftychia Dimitriadou |
E-mail(s) |
eftychia.dimitriadou@med.kuleuven.be
|
Organization name |
KU Leuven
|
Lab |
Cytogenetics and Genome Research
|
Street address |
Herestraat 49, ON1, bus 602
|
City |
Leuven |
ZIP/Postal code |
3000 |
Country |
Belgium |
|
|
Platform ID |
GPL21146 |
Series (2) |
GSE75065 |
Segmental chromosomal imbalances arise at high frequency in human fibroblasts (BAC array) |
GSE75116 |
Segmental chromosomal imbalances arise at high frequency in human fibroblasts |
|