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Sample GSM4094338 Query DataSets for GSM4094338
Status Public on Mar 01, 2021
Title CRC 347 RHAMM- CD44+
Sample type RNA
Source name primary human colorectal cancer
Organism Homo sapiens
Characteristics tissue: colorectal cancer
patient id: 347
cell markers: RHAMM- CD44+
Growth protocol 5000-10000 numbers of cells from freshly isolated colorectal cancer tissues were directly sorted into RNA extraction dilution by FACS.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted using Trizol (Thermo Fisher Scientific; catalog # 15596026), in accordance with the manufacturer’s instructions. RNA was quantified using a NanoDrop-1000 spectrophotometer.
Label Cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
Hybridization protocol 1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
Scan protocol Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
Data processing The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters (protocol GE1-v1_91) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
Submission date Sep 24, 2019
Last update date Mar 02, 2021
Contact name Michitaka Nakano
Organization name Kyushu University
Department Department of Medicine and Biosystemic Sciences
Street address 3-1-1, Maidashi, Higashi-ku
City Fukuoka
State/province Fukuoka
ZIP/Postal code 812-8582
Country Japan
Platform ID GPL17077
Series (1)
GSE137919 RHAMM marks proliferative subpopulation within CD44+ human colorectal cancer stem cells

Data table header descriptions
VALUE Normalized signal intensity

Data table
GE_BrightCorner 6.690846
DarkCorner -6.3100853
A_23_P117082 3.185017
A_33_P3246448 -3.2345858
A_33_P3318220 -6.398809
A_33_P3236322 -4.363018
A_33_P3319925 -3.8904934
A_21_P0000509 9.1350155
A_21_P0000744 -3.2549372
A_24_P215804 -2.2823677
A_23_P110167 4.845835
A_33_P3211513 3.294745
A_23_P103349 -3.0435166
A_32_P61480 -6.3882675
A_33_P3788124 -2.8368497
A_33_P3414202 -1.0202146
A_33_P3316686 0.42643023
A_33_P3300975 -4.22357
A_33_P3263061 3.7860532
A_33_P3261373 -6.018446

Total number of rows: 50739

Table truncated, full table size 1191 Kbytes.

Supplementary file Size Download File type/resource
GSM4094338_SG13284322_253949440299_S001_GE1_1105_Oct12_2_2.txt.gz 3.0 Mb (ftp)(http) TXT
Processed data included within Sample table

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