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Sample GSM4097943 Query DataSets for GSM4097943
Status Public on Sep 26, 2022
Title Hepatocyte_6h_rep2
Sample type RNA
 
Source name Rat primary hepatocyte, 6h, replicate2
Organism Rattus norvegicus
Characteristics cell: primary hepatocyte
gender: male
weight: 320g
Treatment protocol The treatment in our study in the different length of in vitro culture.
Growth protocol Primary hepatocytes isolated from rat liver were resuspended in William's E medium supplemented with 10% FBS (GBICO), 1% ITS, dexamethasone, 1 X L-glutamine/ penicillin /streptomycin and plated onto collagen-I-coated plate at the density of 6× 105 cells/ml. After 3 hours, non-attached cells were washed away and the medium was replaced with William's E free of FBS.
Extracted molecule total RNA
Extraction protocol RNA was prepared using the RNasey Mini Kit (Qiagen p/n 74104) following the manufacturer's recommendations.
Label cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from Quick Amp Labeling Kit, One-Color (Agilent p/n 5190-2305) according to the manufacturer's instructions, followed by RNeasy Mini Kit (Qiagen p/n 74104). Dye incorporation and cRNA yield were checked with theNanoDrop ND-2000 Spectrophotometer.
 
Hybridization protocol 0.6 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 25ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 25ul of 2x GEx Hybridization Buffer was added to the fragmentation mixture and hybridized to Agilent Rat lncRNA Microarray V3 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 Agilent Microarray Scanner (Agilent p/n G2505C) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100%).
Description Gene expression at 6h in replicate2 of primary hepatocytes
Data processing The scanned images were analyzed with Agilent Feature Extraction software using default parameters (protocol GE1-v1_91 and Grid: 012391_D_20060331) 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 26, 2019
Last update date Sep 26, 2022
Contact name jiang zhengyi
E-mail(s) zhengyijiang@zju.edu.cn
Phone 15858111920
Organization name Zhejiang University
Street address Qingchun 79 Road
City Hang Zhou
State/province State...
ZIP/Postal code 310000
Country China
 
Platform ID GPL27537
Series (1)
GSE138071 Integrative proteomics of primary hepatocytes during dedifferentiation

Data table header descriptions
ID_REF
VALUE Normalized signal intensity

Data table
ID_REF VALUE
GE_BrightCorner 15.701753
DarkCorner 2.2009814
A_44_P550748 1.4271911
CUST_R2_06928 5.600013
CUST_R2_17773 1.6769338
A_64_P114354 8.029205
A_44_P307151 2.825382
CUST_R2_03844 9.113815
A_43_P15023 1.6667333
A_44_P1004737 1.730297
A_44_P1088551 1.7181466
CUST_R2_09504 3.777263
CUST_R2_06980 2.580551
CUST_R2_19172 4.2385855
A_64_P119472 1.6432252
CUST_R2_14403 9.226172
CUST_R2_10562 4.246189
A_44_P1096839 4.662482
A_44_P1102733 1.6226804
A_44_P187830 7.820582

Total number of rows: 51323

Table truncated, full table size 1161 Kbytes.




Supplementary file Size Download File type/resource
GSM4097943_liver2-6h.txt.gz 8.4 Mb (ftp)(http) TXT
Processed data included within Sample table

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