|
| Status |
Public on Apr 15, 2015 |
| Title |
Patient 20 NCCRI_2012-054_32 |
| Sample type |
RNA |
| |
|
| Source name |
responder_adjacent normal tissue
|
| Organism |
Homo sapiens |
| Characteristics |
subject id: Patient 20
therapeutic response: responder
gender: Male
age (yrs): 53
tissue: adjacent normal tissue
|
| Treatment protocol |
Twenty-nine patients who received four or more courses of first-line platinum-based chemotherapy after postoperative recurrence were divided into two groups (responder (n=17) and non-responder group (n=12)).
|
| Growth protocol |
Total RNA was extracted from human lungs using QIAzol and the miRNeasy Mini Kit (Qiagen) according to the manufacturer’s protocol. The purity and concentration of all RNA samples were quantified using NanoDrop ND-1000 (Thermo Scientific). Detection of RNA from human lungs was performed using the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA). Prior to the analysis, total RNA was prepared with the Agilent RNA 6000 Pico kit (Agilent Technologies) according to the manufacturer’s protocol. All clinical samples exhibited RNA Integrity Numbers > 6.0.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA samples were extracted from cells using miRNesy mini kit (Quiagen).
|
| Label |
Hy5
|
| Label protocol |
Extracted total RNA was labeled with Hy5 using the miRCURY LNA Array miR labeling kit (Exiqon, Vedbaek, Denmark)
|
| |
|
| Hybridization protocol |
Hybridized for 16 h at 37 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..
|
| Scan protocol |
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
|
| Data processing |
The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals. Measurements of both duplicate spots with the signal intensities greater than 2 standard deviations (SD) of the background signal intensity were considered to be valid. A relative expression level of a given miRNA was calculated by comparing the signal intensities of the averaged valid spots with their mean value throughout the microarray experiments.
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| |
|
| Submission date |
Mar 19, 2014 |
| Last update date |
Apr 15, 2015 |
| Contact name |
Satoshi Kondo |
| Organization name |
Toray Industries,Inc.
|
| Department |
New Projects Development Division
|
| Street address |
Tebiro 6-10-1
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
248-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15446 |
| Series (1) |
| GSE56036 |
microRNA microarray on a cohort study of platinum-sensitive versus platinum-resistant NSCLC patients |
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