|
| Status |
Public on Aug 15, 2024 |
| Title |
ExVivoBlood_patient_3_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Peripheral Blood
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
age: 47y
condition: myasthenia gravis
tissue: PBMCs
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with TRIzol® Reagent (Invitrogen life technologies) from PBMCs following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Each RNA sample was amplified and transcribed into cyanine-3 (Cy3) labeled cRNA along the entire length of the transcripts without 3' bias utilizing a random priming method (Arraystar Flash RNA Labeling Kit, Arraystar). The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000.
|
| |
|
| Hybridization protocol |
1 μg of each Cy3-labeled cRNA was fragmented by adding 5 μl 10 × Blocking Agent and 1 μl of 25 × Fragmentation Buffer, then heated the mixture at 60°C for 30 min, finally 25 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven. After hybridization, microarrays were washed with 1 minute at room temperature GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
lncRNA expression level in PBMCs
|
| Data processing |
Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images.
|
| |
|
| Submission date |
Apr 04, 2024 |
| Last update date |
Aug 15, 2024 |
| Contact name |
Jianjian Wang |
| E-mail(s) |
H06844@hrbmu.edu.cn
|
| Organization name |
The Second Affiliated Hospital of Harbin Medical University
|
| Street address |
No. 246 Xuefu Road, Nangang District, Harbin, China
|
| City |
Harbin |
| ZIP/Postal code |
150081 |
| Country |
China |
| |
|
| Platform ID |
GPL26963 |
| Series (1) |
| GSE263220 |
Analysis of lncRNA expression in myasthenia gravis (MG) and normal subjects |
|
