|
| Status |
Public on Dec 31, 2020 |
| Title |
Diabetic CABG Patient-3 |
| Sample type |
RNA |
| |
|
| Source name |
Pericardial fluid exosome
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Pericardial fluid
cell compartment: exosome
age: 60
gender: Male
nhya class: 2
|
| Treatment protocol |
Samples were used without any further treatment.
|
| Growth protocol |
Exosomes were isolated from the pericardial fluid of the patients undergoing AVR (aortic valve replacement), and CABG (coronary artery bypass graft) surgeries (n=4 in each group) using Exo-SpinTM columns from Cell Guidance Systems (Cambridge, UK). Briefly, 200 µL pericardial fluid was filtered through a sterile 0.22 μm filter (Merck Millipore, Burlington, MA, USA) into a fresh tube. Then, the filtered pericardial fluid was applied onto the Exo-SpinTM columns and exosome separation was performed as described by the manufacturer.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from isolated exosomes using the miRNeasy Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer’s instructions.
|
| Label |
SYBR Green
|
| Label protocol |
A synthetic analog of the non-human Caenorhabditis elegans microRNA-39 (cel-miR-39; Qiagen, Hilden, Germany) was spiked in (10 μl of a 5 fmol/μl stock) to normalize RNA extraction efficiency. cDNA synthesis and real-time qPCR was performed using the miRCURY LNA™ Universal RT microRNA PCR system (Exiqon, Denmark). The cDNA products were transferred to the microRNA PCR Human Panels (I + II; version 4) and run using QuantStudio 6 Flex Real-Time PCR System (Applied Biosystems, Foster City, CA, USA).
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
CABG patient with diabetes
PER128
|
| Data processing |
Immediately after a run,thresholds were determined and data was analyzed using the QuantStudio™ 6 Flex Software. Spike-in control values were checked to determine the accuracy of RNA isolation and cDNA reaction. If a miRNA is expressed in all donors of a group, it was considered as a valid miRNA for the comparison.
No normalization was performed. Raw Different PF exosomes were compared to check whether they express a certain miRNA or not. If Ct<40, we consider the miRNA as present in the exosomes.
The files with non-normalized data on the series record include the well ID.
|
| |
|
| Submission date |
Aug 03, 2018 |
| Last update date |
Jan 01, 2021 |
| Contact name |
Sezin Aday |
| E-mail(s) |
sezin_aday@yahoo.com
|
| Organization name |
University of Pennsylvania
|
| Street address |
210 South 33rd Street, Skirkanich Hall
|
| City |
Philadelphia |
| State/province |
Pennsylvania |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL22600 |
| Series (1) |
| GSE118103 |
Pericardial fluid exosome miRNA profiling |
|
