|
| Status |
Public on Jan 24, 2017 |
| Title |
corneal_endothelium_control_A32 |
| Sample type |
RNA |
| |
|
| Source name |
corneal endothelium control
|
| Organism |
Homo sapiens |
| Characteristics |
gender: Male
age: 37
tissue: corneal endothelium with Descemet's membrane
group: donor cornea as normal control
phakic or pseudophakic: unknown
Cause of death: traffic accident
number of cells per mm²: 2500
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The samples were collected in RNAlater. Tissues were disrupted and homogenized manually with a Disposable Pestle (VWRTM International, Radnor, Pennsylvania). RNA extraction was performed with RNeasy® Micro Kit (Qiagen, Hilden, Germany), without DNase treatment.
|
| Label |
SYBR Green
|
| Label protocol |
PCR assays were performed using two complementary Custom RT² Profiler PCR Arrays (CAPH10409 and CAPH104010, Qiagen) following the manufacturer's instructions. Reverse transcription was performed from 100 ng of total RNA per sample for each 96-well plate, using the RT² First Strand Kit (Qiagen). Quantitative real-time PCR was performed (7900 HT Fast Real-Time PCR System; Applied Biosystems, Carlsbad, California) with 40 cycles at 95°C for 15 seconds and 60°C for 60 seconds, using RT² SYBR Green ROX qPCR Mastermix (Qiagen).
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Control
total RNA (> 200 nucleotides)
|
| Data processing |
Data analysis was done through the web portal of Qiagen, according to the manufacturers instructions: http://qiagen.com/geneglobe. The web-based software automatically performed all ΔΔCt based fold-change calculations from the uploaded raw threshold cycle data. For normalization it used the average arithmetic mean of Ct values from three reference genes: RPL13A, RPL19 and RPS5. To calculate p-values it used a two-sided t-statistic. Sample table reports normalized signal (against housekeeping genes). Fold Change file reports FECD/Control ratios.
|
| |
|
| Submission date |
Dec 03, 2015 |
| Last update date |
Jan 24, 2017 |
| Contact name |
Joost J van den Oord |
| E-mail(s) |
joost.vandenoord@med.kuleuven.be
|
| Phone |
+32 16 33 65 91
|
| Organization name |
KU Leuven
|
| Department |
Department of Imaging & Pathology
|
| Lab |
Translational Cell & Tissue Research
|
| Street address |
Minderbroedersstraat 12, block q, box 1032
|
| City |
Leuven |
| ZIP/Postal code |
3000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL21205 |
| Series (2) |
| GSE75675 |
Identification of Circulating Fibrocytes and Dendritic Derivatives in Corneal Endothelium of Patients with Fuchs' Dystrophy [RT-qPCR array CAPH10410] |
| GSE75676 |
Identification of Circulating Fibrocytes and Dendritic Derivatives in Corneal Endothelium of Patients with Fuchs' Dystrophy |
|
