|
| Status |
Public on Jul 23, 2021 |
| Title |
Post ovulatory phase - participant 15 |
| Sample type |
RNA |
| |
|
| Source name |
Human adipose tissue- post ovulatory phase
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Subcutaneous adipose
age: 29
bmi: 22.72
menstrual cycle phase: Post ovulatory
menstrual cycle day: 17
|
| Treatment protocol |
Immediately following the biposy, adipose tissue samples were placed into an RNA-free 1.5 ml microcentrifuge tube containing 5-10 x tissue volume RNAlater (ThermoFisher, UK, cat no. AM7020), flash frozen in liquid nitrogen and stored at -80 ¬oC until analysis.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from 30-100 mg homogenised adipose tissue in a combined protocol using TRIzol reagent (ThermoFisher Scientific, UK, cat no. 15596026) and miRNeasy mini lipid kit (Qiagen, Germany, cat no. 217004), as previously described by Cirera et al. 2013 (doi:10.1186/1756-0500-6-472).
|
| Label |
Biotin
|
| Label protocol |
100 ng RNA were labelled using the Flashtag Biotin HSR labelling kit according to manufacturers instructions
|
| |
|
| Hybridization protocol |
Hybridisation cocktails were prepared using the GeneChip hybridisation kit (ThermoFisher Scientific, UK, cat no. 900720). Biotin labelled samples were hybridised overnight onto GeneChip miRNA 4.0 arrays (ThermoFisher Scientific, UK, cat no. 902413) according to manufacturer’s instructions. Arrays were washed and stained using the GeneChip wash and stain kit (ThermoFisher, UK, cat no. 902413) and the Affymetrix fluidics station 450, as per manufacturer’s instructions.
|
| Scan protocol |
GeneChips were scanned by the Affymetrix scanner 3000 7G.
|
| Description |
Adipose tissue miRNA expression from the post ovulatory phase
|
| Data processing |
Microarray signal intensities were normalised using the variance stabilisation normalisation 2 (VSN2) algorithm and probe level summarisation conducted using robust multi-chip analysis (RMA).
|
| |
|
| Submission date |
Jul 22, 2021 |
| Last update date |
Jul 23, 2021 |
| Contact name |
Colin N Moran |
| Organization name |
University of Stirling
|
| Lab |
Physiology, Exercise and Nutrition Research Group
|
| Street address |
Univeristy of Stirling
|
| City |
Stirling |
| ZIP/Postal code |
FK9 4LA |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL19117 |
| Series (1) |
| GSE180625 |
Changes in adipose tissue microRNA expression across the menstrual cycle in regularly menstruating women: a pilot study. |
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