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Sample GSM429635 Query DataSets for GSM429635
Status Public on Feb 04, 2011
Title DG75-eGFP Ago2-IP; 1. Replicate
Sample type RNA
 
Source name DG75-eGFP Ago2-IP
Organism Homo sapiens
Characteristics antibody: Ago2
replicate: 1
Growth protocol DG75-eGFP, DG75-10/12 and BCBL-1 were cultured in RPMI 1640 medium containing 100 IU/ml Penicillin, 100 µg/ml Streptomycin and 2 mM L-Glutamin. BL41, BL41 B95.8 and Jijoye were cultered in RPMI 1640 medium containing 10% fetal calf serum and Penicillin/Streptomycin as well as Bykomycine and Nystatin.
Extracted molecule total RNA
Extraction protocol 5x10e08 cells were lyzed in 10 ml of lysis buffer containing 25 mM Tris HCl pH 7.5, 150 mM KCl, 2 mM EDTA, 0.5% NP-40, 0.5 mM DTT, and protease inhibitors (Roche); incubated for 30 min at 4°C and cleared by centrifugation at 20,000 g for 30 min. Conditions for immunoprecipitation: 60 µl Protein-G sepharose beads were incubated with 5 m RPMI containing 6µg of either anti hAgo2 (11A9) or anti-BrdU (abcam) over night. Next morning columns were drained by gravity flow and washed once with lysis buffer. Next, 5 ml of lysates were added and incubated for 3 h at 4°C. In the meanwhile, total RNA was prepared from 100µl of cell lysates. This was analyzed for BL41, B41 B95.8 and Jijoye instead of the BrdU-IP. Following the incubation, beads were washed 4x with IP wash buffer (300 mM NaCl, 50 mM Tris HCl pH 7.5, 5 mM Mg2Cl, 0.1% NP-40, 1 mM NaF) and once with PBS. RNA was recovered using miRNeasy (Qiagen).
Label biotin
Label protocol Affymetrix protocol
 
Hybridization protocol Affymetrix protocol
Scan protocol Affymetrix protocol
Description gene expression data
Data processing RMA
 
Submission date Jul 17, 2009
Last update date Oct 29, 2013
Contact name Lars Dölken
E-mail(s) Lars.Doelken@uni-wuerzburg.de
Organization name University of Cambridge
Department Medicine
Lab Box 157, Level 5
Street address Addenbrookes Hospital, Hills Road
City Cambridge
State/province Cambridgeshire
ZIP/Postal code CB20QQ
Country United Kingdom
 
Platform ID GPL6244
Series (1)
GSE17180 Identification of KSHV, EBV and cellular miRNA targets in human B-cells using RIP-Chip

Data table header descriptions
ID_REF
VALUE RMA signal intensity

Data table
ID_REF VALUE
8064336 1046.6
8117020 983.8
7973618 417.3
8130539 883.5
7982868 1543.0
8115865 1591.4
8036908 492.8
8066716 702.7
7963009 100.2
7908917 3286.7
8005699 323.4
8018324 515.8
8029560 2175.9
7911559 1501.2
8039939 1501.2
7947434 1033.8
7905918 537.6
8177955 842.0
8052834 632.4
8118116 1128.3

Total number of rows: 33297

Table truncated, full table size 420 Kbytes.




Supplementary file Size Download File type/resource
GSM429635.CEL.gz 4.0 Mb (ftp)(http) CEL
Processed data included within Sample table

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