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Status |
Public on Jan 01, 2012 |
Title |
IDD patient 3 |
Sample type |
RNA |
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|
Source name |
Nucleus pulposus cells, degenerative
|
Organism |
Homo sapiens |
Characteristics |
tissue: Nucleus pulposus condition: degenerative disease: intervertebral disc degeneration (IDD) gender: female age: 33 degeneration grading: Grading IV
|
Extracted molecule |
total RNA |
Extraction protocol |
Nucleus pulposus (NP) cells were isolated by enzymatic digestion of collected human NP for 40 min at 37 °C in PBS with 0.25% pronase (Gibco-BRL) and subsequently for 4 h in PBS with 0.025% collagenase type II (Invitrogen). Total RNA of NP cells was extracted using Trizol(Invitrogen) and RNeasy mini kit (QIAGEN) according to manufacturer’s instructions.
|
Label |
Hy3
|
Label protocol |
The samples were labeled using the miRCURY™ Hy3™/Hy5™ Power labeling kit (Exiqon) according to manufacturer's instructions.
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|
|
Hybridization protocol |
The labeled samples were hybridized on the miRCURY LNA™ microRNA Array V11.0.
|
Scan protocol |
Hybridization signals were detected by fluorescence using an Axon GenePix 4000B microarray scanner. Scanner images were quantified by GenePix Pro 6 (Molecular Devices).
|
Description |
Biological replicate 3 of 3. Degenerative human nucleus pulposus cells harvested following digestion of nucleus pulposus with pronase and collagenase II.
WDN
|
Data processing |
The intensity of green signal was calculated after background subtraction and the median value of four replicated spots of each probe on the same slide had been calculated. We used Median Normalization Method to obtain “Normalized Data”, Normalized Data=(Foreground-Background)/median, the median is 50 percent quantile of microRNA intensity which is larger than 50 in all samples after background correction.
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|
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Submission date |
Jan 19, 2010 |
Last update date |
Feb 20, 2014 |
Contact name |
Hai-Qiang Wang |
E-mail(s) |
hqwang@sntcm.edu.cn, drwanghq@163.com
|
Organization name |
Shaanxi University of Chinese Medicine
|
Department |
Insititute of Integrative Medicine
|
Street address |
Xixian Avenue, Xixian District
|
City |
Xi'an |
ZIP/Postal code |
712046 |
Country |
China |
|
|
Platform ID |
GPL9946 |
Series (2) |
GSE19943 |
MicroRNAs expression profiling of human nucleus pulposus cells: control vs. degeneration |
GSE67567 |
Noncoding RNAs in human intervertebral disc degeneration: an integrated microarray study |
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