|
| Status |
Public on Sep 04, 2018 |
| Title |
thymus_non-puberty_NF1_miRNA |
| Sample type |
RNA |
| |
|
| Source name |
thymus
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: thymus
gender: female
age at surgery (yr/mo/d): 1yr5mo
|
| Treatment protocol |
Fresh ex-vivo explants of patients were obtained at the surgery room and immediately preserved with 600 uL of RNAlater (Qiagen).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Thymus explants (3-4 mm3) were homogenized with TissueRupter (Qiagen) and total RNA was extracted from the homogenates using the RNeasy Lipid Tissue Kit (Qiagen)
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.05 ug RNA using the microRNA Complete Labeling and Hyb Kit (Agilent Technologies), without column purification, according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Dried Cy3-labelled miRNA sample was resuspended with 17uL of nuclease-free water and mixed to Hyb Spike-In and 10x GE Blocking Agent solutions, and 2x Hi-RPM Hybridization buffer. The final volume was incubated at 100°C for 5 minutes and hybridized to Human miRNA Microarray slides (8x60K) - (G4872A) - Agilent Technologies, for 20 hours at 55°C in a rotating Agilent hybridization oven (20rpm).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent Bundle using one color scan setting for 8x60k miRNA microarrray slides.
|
| Description |
microRNA expression from thymus explant
|
| Data processing |
Data extraction was carried out with Feature Extraction Software 10.7.3 (Agilent) and considering spots present none or only one flag (i.e. low intensity, saturation, controls, etc.). The selected miRNA transcripts were used for analysis using the R software and considering up to 3 NA for each group (MM or MF) and the expression values in Log (base 2). NA converted to null in data table. Data for identical probe names were averaged. Data not normalized.
|
| |
|
| Submission date |
Apr 24, 2018 |
| Last update date |
Sep 04, 2018 |
| Contact name |
Carlos Alberto Moreira-Filho |
| E-mail(s) |
[email protected]
|
| Phone |
55-11-3061-8449
|
| Organization name |
Faculdade de Medicina da Universidade de Sao Paulo
|
| Department |
Departments of Pediatrics
|
| Lab |
LIM36
|
| Street address |
Av. Dr. Eneas Carvalho de Aguiar 647
|
| City |
Sao Paulo |
| State/province |
SP |
| ZIP/Postal code |
05403-900 |
| Country |
Brazil |
| |
|
| Platform ID |
GPL21576 |
| Series (2) |
| GSE113596 |
Minipuberty and Thymic Sexual Dimorphism in the Infant Human Thymus [non-puberty miRNA] |
| GSE113597 |
Minipuberty and Sexual Dimorphism in the Infant Human Thymus |
|
