|
| Status |
Public on Apr 01, 2024 |
| Title |
CRF433 |
| Sample type |
RNA |
| |
|
| Source name |
Human peripheral blood
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Peripheral blood
cell type: Monocytes
participant group: Non-pregnant
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Sample processing: 1 x 106 of monocytes were washed with 1 ml RPMI 1640/Glutamax and centrifuged at 8,000 x g for 4 mins. The cells were re-suspended with diluted RLT buffer (Qiagen, UK) (1/3 with RNase-free water; Invitrogen) to 10,000 cells / μl, and frozen at -80°C.
|
| Label |
n.a.
|
| Label protocol |
Sample RNA is hybridised with target-specific capture and reporter probes in order to create a library of unique target-probe complexes. Each target is labelled with a specific molecular barcode, which can then be detected and counted using an automated fluorescence microscope
|
| |
|
| Hybridization protocol |
RNA samples stored at -80°C were thawed on ice prior to hybridisation reactions. Reporter probes were diluted in 70 μl of hybridisation buffer and gently mixed to create a master mix. To set up the hybridisation reactions, 5 μl of each sample was added to 8 μl of master mix. Hybridisation reactions were then completed by adding 2 μl of Capture ProbeSet and incubation at 65°C for 20 h. Probe set-target RNA hybridisation solution is then further diluted to a final volume of 35 μl. 33 μl of the diluted hybridisation solution is loaded onto a SPRINT microfluidic cartridge, with an air bubble also inserted to the stop point.
|
| Scan protocol |
Detection and scanning were performed by the nCounter® SPRINT Profiler analysis system (Nanostring).
|
| Description |
CRF433
|
| Data processing |
Data analysis was performed using the advanced analysis package (2.0.134) within the nSolver© analysis software (Version 4.0, NanoString©), and differential expression p-value adjustment was done with Benjamini-Hochberg. Significant gene changes were determined to be p ≤ 0.05 with a fold change of ≤ -0.5 or ≥0.5.
|
| |
|
| Submission date |
Feb 07, 2024 |
| Last update date |
Apr 01, 2024 |
| Contact name |
April Rees |
| E-mail(s) |
april.rees@swansea.ac.uk
|
| Phone |
01792987806
|
| Organization name |
Swansea University
|
| Street address |
Room 203 ILS1, Singleton Park
|
| City |
Swansea |
| ZIP/Postal code |
SA2 8PP |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL33903 |
| Series (1) |
| GSE255271 |
Immunometabolic adaptation in monocytes underpins functional changes during pregnancy [metabolism] |
|
