|
| Status |
Public on Nov 06, 2018 |
| Title |
WBS_081 |
| Sample type |
RNA |
| |
|
| Source name |
Peripheral blood from WBS patient
|
| Organism |
Homo sapiens |
| Characteristics |
diagnosis: Williams Syndrome (WS)
tissue: whole blood
age: 24y
gender: female
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNA was extracted from peripheral blood of participants by using the PAXgene Blood miRNA System (QIAGEN, Tokyo, Japan) following the manufacturer's protocol with DNase treatment step. RNA was quantified using a NanoDrop-2000 spectrophotometer (Thermo Fisher Scientific, Yokohama, Japan), and quality was determined by the Agilent 2100 Bioanalyzer (Agilent Technologies, Tokyo, Japan).
|
| Label |
Cy3
|
| Label protocol |
Following manufacturer's instructions, total RNA (100ng) was labeled with Cy3 using the miRNA Complete Labeling and Hyb Kit(Agilent Technologies, Tokyo, Japan).
|
| |
|
| Hybridization protocol |
Following manufacturer's instructions, labelled RNA was hybridized to the SurePrint G3 Human miRNA 8x60K Microarray (G4870C) (Agilent Technologies) using the miRNA Complete Labeling and Hyb Kit (Agilent Technologies). Hybridization step was performed for 20 hours at 55°C at a constant speed rotation of 20rpm. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent Technologies) and 5 minute with 37°C GE Wash buffer 2 (Agilent Technologies), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on Agilent Microarray Scanner(p/n G2565BA) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 3um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.0.1.1 (Agilent Technologies) using default parameters (protocol: miRNA_1100_Jul11 and Grid: 070156_D_F_20141006) to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Nov 07, 2016 |
| Last update date |
Nov 06, 2018 |
| Contact name |
Ryo Kimura |
| E-mail(s) |
[email protected]
|
| Organization name |
Kyoto University Graduate School of Medicine
|
| Department |
Anatomy and Developmental Biology
|
| Street address |
Yoshida-Konoe-cho
|
| City |
Kyoto |
| ZIP/Postal code |
606-8501 |
| Country |
Japan |
| |
|
| Platform ID |
GPL21575 |
| Series (1) |
| GSE89595 |
Signiture microRNA expression patterns identified in Williams syndrome |
|
