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Sample GSM2747732 Query DataSets for GSM2747732
Status Public on Aug 01, 2019
Title B16F0 cells cultured for 24 hr in serum-free media, rep 3
Sample type RNA
Source name B16F0 cells
Organism Mus musculus
Characteristics tag: B16F0
Treatment protocol not applicable
Growth protocol Cultured for 24 hours in serum-free media
Extracted molecule total RNA
Extraction protocol Total RNA isolated from B16F0 exosomes and cells by RNeasy kit (Qiagen) was quantified using Nanodrop and analyzed by on-chip-electrophoresis using the Agilent Bioanalyzer.
Label biotin
Label protocol Four hundred nanograms of each miRNA sample was biotin labeled using the Genisphere FlashTag HSR Kit according to the manufacturer's instructions. The labeling of RNA was confirmed by Enzyme Linked Oligosorbent Assay (ELOSA) according Genisphere FlashTag HSR protocol.
Hybridization protocol The 21.5 microl of biotin-labeled RNA with added hybridization controls was hybridized to the GeneChip miRNA 2.0 Arrays (Affymetrix) at 48oC and 60 rpm for 16 hours in GeneChip Hybridization Oven 640 (Affymetrix).The entire reaction of fragmented and biotin-labeled cDNA (50 mu) with added hybridization controls was hybridized to the mouse GeneChip 1.0 ST Exon Arrays (Affymetrix) at 45oC for 17 hours in GeneChip Hybridization Oven 640 (Affymetrix).
Scan protocol GeneChip miRNA 2.0 Arrays were stained using FS 450_0003 protocol in Affymetrix GeneChip Fluidics Station 450. Briefly, biotin-labeled RNA was reacted using washes with a solution containing a streptavidin-phycoerythrin complex, with an intermediate treatment of biotin-labeled anti-streptadvidin antibody to amplify the signal. Phycoerythrin labeling was detected within the Affymetrix GeneChip Scanner 3000 7G plus using 532 nm light and detected by a photomultiplier tube.
Data processing A miRNA QC Tool software (Affymetrix) was used to check quality controls of hybridized chips. All chips that passed quality controls were RMA normalized using miRNA QC Tool.
Submission date Aug 21, 2017
Last update date Aug 01, 2019
Contact name David John Klinke
Phone 304-293-9346
Organization name West Virginia University
Department Chemical Engineering
Street address P.O. Box 6102
City Morgantown
State/province WV
ZIP/Postal code 26506-6102
Country USA
Platform ID GPL14613
Series (2)
GSE102883 Expression data of miRNA from parental B16F0 cells and B16F0 exosomes [miRNA]
GSE102951 Expression data from parental B16F0 cells and B16F0 exosomes

Data table header descriptions
VALUE RMA signal estimates from Expression Console Software

Data table
cel-let-7_st 12.1796 8.72E-16
cel-let-7-star_st 3.897316 0.9468079
cel-lin-4_st 3.998224 0.4228857
cel-lin-4-star_st 3.966379 0.5989928
cel-miR-1_st 3.949857 0.3601989
cel-miR-2_st 3.904985 0.9049826
cel-miR-34_st 3.910461 0.8990512
cel-miR-34-star_st 4.223197 0.1145662
cel-miR-35_st 4.040006 0.6351479
cel-miR-36_st 4.063942 0.02849079
cel-miR-37-star_st 4.087666 0.6435205
cel-miR-37_st 3.691853 0.972202
cel-miR-38_st 4.08014 0.5555594
cel-miR-39_st 3.85409 0.8330896
cel-miR-40_st 4.059304 0.2395055
cel-miR-41_st 3.932922 0.958921
cel-miR-42-star_st 3.914288 0.8638405
cel-miR-42_st 3.914288 0.9031236
cel-miR-43_st 3.792145 0.9902807
cel-miR-44_st 4.076582 0.8555715

Total number of rows: 20180

Table truncated, full table size 701 Kbytes.

Supplementary file Size Download File type/resource
GSM2747732_3_cell_24h_miRNA-2_0_.CEL.gz 627.7 Kb (ftp)(http) CEL
Processed data included within Sample table

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