|
| Status |
Public on Jun 10, 2023 |
| Title |
256740610807_1_4,Sd11,hacontrol |
| Sample type |
RNA |
| |
|
| Source name |
lymphocytes, Kazakh, healthy control
|
| Organism |
Homo sapiens |
| Characteristics |
population: Xinjiang Kazakh
diagnosis: healthy
cell type: peripheral blood lymphocytes
gender: male
|
| Extracted molecule |
total RNA |
| Extraction protocol |
5µL Total RNA (100-500 ng) was taken and added to 0.2 mL nuclease-free centrifuge tube.
All samples were added to Spike A (Agilent) 2.0µL or Spike B (Agilent) 2.0µL.
Prepare reverse transcription Master Mix on ice, mix gently, centrifuge briefly and place on ice bath.
5µL of reverse transcription Master Mix was added to a 0.2mL centrifuge tube containing the Total RNA sample. Reverse transcription
The final reaction volume is 10µL.
Gently blow and mix for 2-3 times, centrifuge instantaneously, and place on ice.
The reverse transcription centrifuge tube was placed on the PCR apparatus, and the reaction time was 1h at 25℃, 1h at 42℃, and kept at 4℃ for more than 5min.
The reverse transcription centrifuge tube was removed, centrifuged instantaneously and placed on ice to prepare for the Second Strand cDNA synthesis reaction.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturer's instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565CA) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Sd11
The expression level of lncRNA in peripheral blood lymphocytes of Kazakh healthy participants was detected by gene chip technology.
|
| Data processing |
Feature Extraction software was used to preprocess tiff image data, and GeneSpring GX software was used to calculate gene expression difference and statistical significance p value. The tiff image is scanned on the chip and FeatureExtraction software is used to obtain a raw data file (.txt).mport the original data file (.txt) into GeneSpring software and write the grouping and other parameter information. Data normalization and QC analysis were performed for each sample. Cluster3.0 software is used for Cluster analysis and graphical presentation.
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| |
|
| Submission date |
Jun 05, 2023 |
| Last update date |
Jun 10, 2023 |
| Contact name |
Tongjian Zhao |
| Organization name |
School of Pharmaceutical Sciences, Jilin University
|
| Street address |
1266 Fujin Road
|
| City |
Changchun |
| ZIP/Postal code |
130021 |
| Country |
China |
| |
|
| Platform ID |
GPL20115 |
| Series (1) |
| GSE234085 |
Differential expression of lncRNA in peripheral blood lymphocytes of Xinjiang Kazakh patients with essential hypertension |
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