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Status |
Public on Jan 24, 2017 |
Title |
corneal_endothelium_control_A62 |
Sample type |
RNA |
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Source name |
corneal endothelium control
|
Organism |
Homo sapiens |
Characteristics |
gender: Female age: 72 tissue: corneal endothelium with Descemet's membrane group: donor cornea as normal control phakic or pseudophakic: unknown Cause of death: edema ascites decompensation number of cells per mm²: 2400
|
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).
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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.
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|
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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 |
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