|
| Status |
Public on Jul 28, 2021 |
| Title |
CUTO44 |
| Sample type |
genomic |
| |
|
| Source name |
Patient-derived lung adenocarcinoma cell line
|
| Organism |
Homo sapiens |
| Characteristics |
gender: female
tissue: Liver
disease state: Lung adenocarcinoma
|
| Treatment protocol |
No treatement were done on cell pellet
|
| Growth protocol |
CUTO44 cell line were cultured in RPMI-1640 with 10%FBS and grown on a 10cm TC Dish. When 100% confluency was reach, cells were trypsinized and spun down as cell pellet
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Genomic DNA was isolated using a BioRobot M48 Workstation with MagAttract technology (Qiagen)
|
| Label |
A-DNP, C-Bio
|
| Label protocol |
The MSA1 plate was prepared according to Illuminia protocol. 200ng of gDNA was added to the wells and denatured with 4ul of 0.1NaOH for 10mins at room temperature(RT). 68ul of random primer mix and 75ulmulti-sample ampification master mix was added to the well. The plate was place in the Illumina Hyb oven for 24hours at 37C for uniform amplification. The sample then underwent enzymatic DNA fragmentation for 1 hour at 37C followed by DNA precipitation with 100% isopropanol and PM1 solution followed by air drying the MSA1 plate for 1 hour. Prior to hybridization, the dry sample was resuspended in RA1 buffer
|
| |
|
| Hybridization protocol |
The resuspended, fragmented DNA sample was denature at 95C for 20mins, cooled for 30mins and then loaded onto a prepared BeadChip and place in the Hyb Chamber for 24hr hybrization at 48C. The BeadChip was wash with PB1 to prep it for Xstain and single-base extension follow by Bead Chip staining. Finally, the BeadChip was washed with PB1 and coated with XC4/ethanol mix. BeadChip was dried till scanning.
|
| Scan protocol |
The dried BeadChip was imaged using the iScan Reader according to illumina protocol
|
| Description |
Infinium HumanCytoSNP-850K v2.1 BeadChips
|
| Data processing |
Genomic DNA extracted from patient-derived LUAD cell line was genotyped using Infinium HumanCytoSNP-850K-12 v2.1 BeadChips (Illumina).
|
| |
|
| Submission date |
Jul 27, 2021 |
| Last update date |
Jul 28, 2021 |
| Contact name |
Robert Doebele |
| E-mail(s) |
anh.le@cuanschutz.edu
|
| Phone |
3037240033
|
| Organization name |
University of Colorado-AMC
|
| Department |
Medical Oncology
|
| Lab |
Doebele Lab
|
| Street address |
12801 E. 17th Ave
|
| City |
AURORA |
| State/province |
CO |
| ZIP/Postal code |
80045 |
| Country |
USA |
| |
|
| Platform ID |
GPL19718 |
| Series (1) |
| GSE180894 |
The evolution of MET and NRAS gene amplification as sequential acquired resistance mechanisms in a patient with EGFR mutation positive non-small cell lung cancer |
|
