|
| Status |
Public on Apr 01, 2014 |
| Title |
CRC3a_serum_exo_8_after |
| Sample type |
RNA |
| |
|
| Source name |
Colorectal cancer patient, after surgical resection, TNM Stage IIIa, serum, exosomes,8
|
| Organism |
Homo sapiens |
| Characteristics |
microrna source: exosomes
biological source of exosomes: serum
gender: female
age: 55
disease state: Colorectal cancer
tnm stages: IIIa
time: after surgical resection of primary tumor
|
| Treatment protocol |
Peripheral blood sera from colorectal cancer patients were diluted 1:7 with PBS. Exosomes fraction were prepared by step-wise centrifugation-ultracentrifugation method.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Exosome fraction was mixed with Trizol-LS reagent (Invitrogen), and aqueous phase was collected by adding chloroform. After addition of ethanol to the aqueous phase, it was placed on to an RNeasy mini spin column (Qiagen) for the purification of total RNAs.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled miRNA was prepared from total RNA using the miRNA Complete Labeling and Hyb Kit (Agilent)and MicroRNA Spike-In Kit (Agilent) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Cy3-labelled miRNA was incubated at 100°C for 5 minutes in a reaction volume of 45uL containing Hyb Spike-In, 2x Hi-RPM Hybridization Buffer and 10x Gene Expression blocking agent following the manufacturers instructions and cooled down in ice-cold water for 5 min. Reaction mixture hybridized to Agilent Human miRNA V3 Microarray (G4470C) for 20 hours at 55°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent) and 5 minute with 37°C GE Wash buffer 2 (Agilent).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x15k array slides (Scan Area 61x21.6 mm, Scan resolution 5 um, Dye channel is set to Green ).
|
| Description |
MicroRNA profile in serum exosomes of colorectal cancer patients after sergical resection of primary tumor
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1 (Agilent) using default parameters (protocol miRNA_107_Sep09) and Grid: 021827_D_F_20081129) to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Aug 21, 2012 |
| Last update date |
Apr 01, 2014 |
| Contact name |
Naoto Tsuchiya |
| Organization name |
National Cancer Center Reasearch Institute
|
| Lab |
Genome Biology
|
| Street address |
Tsukiji 5-1-1
|
| City |
Chuo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
104-0045 |
| Country |
Japan |
| |
|
| Platform ID |
GPL14767 |
| Series (2) |
| GSE40246 |
microRNA profiles of serum exosomes in healthy controls and colorectal cancer patients-2 |
| GSE40247 |
Profiling of endogenous and exosomal microRNAs in colon cancer |
|
