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Sample GSM2747737 Query DataSets for GSM2747737
Status Public on Aug 01, 2019
Title B16F0 exosomes isolated from serum-free media media conditioned by B16F0 cells for 24 hr, rep 4
Sample type RNA
Source name B16F0 exosomes
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.
Description exosomal RNA
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 11.56026 8.72E-16
cel-let-7-star_st 3.739862 0.9866574
cel-lin-4_st 4.157996 0.07223933
cel-lin-4-star_st 4.119008 0.6356521
cel-miR-1_st 4.11658 0.6045325
cel-miR-2_st 4.349701 0.2648486
cel-miR-34_st 3.973235 0.6585042
cel-miR-34-star_st 7.219577 1.97E-05
cel-miR-35_st 3.779269 0.8003891
cel-miR-36_st 4.114903 0.1670159
cel-miR-37-star_st 3.995642 0.7804124
cel-miR-37_st 3.946352 0.7773904
cel-miR-38_st 3.946945 0.9069493
cel-miR-39_st 4.157518 0.6375713
cel-miR-40_st 4.316307 0.2219788
cel-miR-41_st 4.115423 0.5070394
cel-miR-42-star_st 3.639858 0.9557202
cel-miR-42_st 3.877966 0.9423763
cel-miR-43_st 3.762909 0.9301788
cel-miR-44_st 3.940183 0.7951032

Total number of rows: 20180

Table truncated, full table size 703 Kbytes.

Supplementary file Size Download File type/resource
GSM2747737_8_exosome_24h_miRNA-2_0_.CEL.gz 542.2 Kb (ftp)(http) CEL
Processed data included within Sample table

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