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Sample GSM3318619 Query DataSets for GSM3318619
Status Public on Dec 31, 2020
Title Diabetic CABG Patient-4
Sample type RNA
 
Source name Pericardial fluid exosome
Organism Homo sapiens
Characteristics tissue: Pericardial fluid
cell compartment: exosome
age: 75
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
PER129
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

Supplementary file Size Download File type/resource
GSM3318619_2017-01-02_PER129_Panel_1.xls.gz 2.1 Mb (ftp)(http) XLS
GSM3318619_2017-01-09_PER129_Panel_2.xls.gz 2.1 Mb (ftp)(http) XLS
Processed data not provided for this record

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