|
| Status |
Public on Aug 24, 2021 |
| Title |
PC9 siYTHDF2 #2 |
| Sample type |
RNA |
| |
|
| Source name |
siYTHDF2 #2
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: PC9
cell type: lung adenocarcinoma
genotype/variation: siYTHDF2
|
| Treatment protocol |
Cells were cultured for 24 hours prior to transfection and transfected with 15 nM of final concentrations of siRNAs. Total RNA were extracted at 48 h after transfection.
|
| Growth protocol |
Cells were maintained in a 5% CO2 and 95% air incubator at 37℃.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy Plus Mini Kit (#74136, Qiagen) according to the manufacturer’s instructions.
|
| Label |
NanoString Oligonucleotide Probe
|
| Label protocol |
The RNA samples were run on the NanoString nCounter Analysis System (NanoStringTechnologies, Inc., WA, USA) according to the manufacturer's directions.
|
| |
|
| Hybridization protocol |
5μL (100-300ng) of each RNA sample was mixed with 8μL of the Master Mix (reporter codeSet and hybridization buffer). 2μL of the capture probeSet was added; the solution was mixed and spun down. It was placed in a 65°C thermocycler (Bio-Rad Laboratories Inc, Hercules, California, USA) for 16 hours (maximum hybridization time should not exceed 48 hours).
|
| Scan protocol |
The samples were transferred to the preparation station (NanoString Technologies, Seattle, WA) with prepared nCounter Master Kit and a cartridge. The samples binding to the cartridge. The preparation station 12 lanes per run in approximately 2.5 to 3 hours. The cartridges were transferred to the Digital Analyzer (NanoString Technologies, Seattle, WA) for analysis. Cartridges were then scanned on the Digital Analyzer at 555 fields of view.
|
| Description |
Gene expression data from PC9 cells
|
| Data processing |
To quantify gene expression values, first normalization step was analyzed by geNormalgorithm in nCounter Advanced Analysis ver2.0.115.
|
| |
|
| Submission date |
Apr 07, 2021 |
| Last update date |
Aug 24, 2021 |
| Contact name |
Kazuo Tsuchiya |
| E-mail(s) |
bandfor2004@yahoo.co.jp
|
| Organization name |
Hamamatsu University School of Medicine
|
| Department |
Tumor pathology
|
| Street address |
handayama1-20-1
|
| City |
Hamamatsu |
| ZIP/Postal code |
4313192 |
| Country |
Japan |
| |
|
| Platform ID |
GPL27956 |
| Series (1) |
| GSE171634 |
Expression data from YTHDF1 or YTHDF2 down-regulated PC9 |
|
