|
| Status |
Public on Aug 29, 2020 |
| Title |
Series 3_KD |
| Sample type |
RNA |
| |
|
| Source name |
HEK-293FT
|
| Organism |
Homo sapiens |
| Characteristics |
genotype: TruB1 KD
cell line: HEK-293FT
|
| Treatment protocol |
siRNAs were transfected into cells using Lipofectamine RNAiMAX (Thermofisher scientific) by manufacturer’s recommended protocol.
|
| Growth protocol |
HEK293FT cells were maintained in DMEM medium (Corning) supplemented with 10% FBS (Gibco) and 1% penicillin–streptomycin (Gibco) at 37 °C with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by mirVana miRNA isolation kit.
|
| Label |
TaqMan
|
| Label protocol |
3μL of RNA (1000 ng) were reverse transcribed by Megaplex RT Primers, Pool A (human) (Thermo Fisher Scientific) using the TaqMan® microRNA Reverse Transcription Kit (Thermo Fisher Scientific). For qRT-PCR, RT product was mixed with TaqMan® Universal PCR Master Mix, No AmpErase UNG (Thermo Fisher Scientific) and the appropriate amount of water, and subsequently loaded into the ports of the TaqMan array cards. RealTime PCR was performed on a 7900HT Fast Real-Time PCR System with TaqMan® Array Block (Thermo Fisher Scientific), using universal cycling conditions (95 °C/10 min, then [95 °C/15 sec, 60 °C/60 sec] for 40 cycles).
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
test
SAMPLE 6
|
| Data processing |
For the normalization it uses the average of five housekeeping genes: RNU6B. Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_RNU6B)]
processed_data.txt normalized signal (against housekeeping genes).
Fold-change.txt reports test/control (KD/control) ratios.
|
| |
|
| Submission date |
Jan 13, 2020 |
| Last update date |
Aug 29, 2020 |
| Contact name |
Ryota Kurimoto |
| Organization name |
Tokyo Medical and Dental University
|
| Department |
Systems BioMedicine
|
| Street address |
1-7-5
|
| City |
Yushima, Bunkyo-ku, Tokyo |
| State/province |
Tokyo |
| ZIP/Postal code |
113-8519 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15634 |
| Series (1) |
| GSE143508 |
Real-time quantitative PCR analysis of TruB1 KD HEK-293FTcells |
|
