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Status |
Public on Nov 08, 2013 |
Title |
IP Flag HeLa 9 18.03 |
Sample type |
RNA |
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Source name |
HeLa_control IP
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Organism |
Homo sapiens |
Characteristics |
cell line: HeLa H9 ip antibody: anti-Flag Antibody antibody vendor: Sigma antibody cat. #: F3165 antibody lot #: SLBF1225
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Treatment protocol |
Cells were extracted in HNTG buffer (20 mM HEPES, pH 7.9, 150 mM NaCl, 1% Triton, 10% glycerol, 1 mM MgCl2, 1 mM EGTA, and protease inhibitors) for 30 min at 4°C. Cellular debris were removed by centrifugation (10 min at 9000 g). Extracts were put on antibody-coated beads for 2h at 4°C (M2 -Sigma). Beads were washed twice in HNTG and three times in PBS, and pelleted materials were Trizol.
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Growth protocol |
HeLa cells were grown to sub-confluency in DMEM, 10% FCS, at 37°C.
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Extracted molecule |
total RNA |
Extraction protocol |
RNAs from beads or cell extracts were homogenized in Trizol and purified with RNeasy mini kit (Qiagen).
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Label |
biotin
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Label protocol |
Samples were prepared and hybridized according to Affymetrix specifications. (Tilling with WT Amplified Double-Stranded cDNA Synthesis Kit)
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Hybridization protocol |
Samples were prepared and hybridized according to Affymetrix specifications. (kit Hybridization Wash and Stain kit réf: 900720)
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Scan protocol |
The tiling arrays were scanned using an Affymetrix GeneChip Scanner 3000 7G according to manufacturer's instruction.
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Description |
result file: Ars2VsHeLa_signal.txt; IPCbpVsIPHela_signal.txt
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Data processing |
The signal intensity data was analyzed with the Affymetrix Tiling Analysis Software (v. 1.1) using parameters: one side upper, 90 bp bandwidth and perfect match only. Output txt files (i.e. result file). TASParam* files include parameter settings and input file information for the TAS analysis for each result file.
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Submission date |
Nov 06, 2013 |
Last update date |
Nov 08, 2013 |
Contact name |
Edouard BERTRAND |
Organization name |
CNRS
|
Street address |
1919 Route de Mendes
|
City |
Montpellier |
ZIP/Postal code |
34293 |
Country |
France |
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Platform ID |
GPL4910 |
Series (1) |
GSE52132 |
The human cap-binding complex is functionally connected to the nuclear RNA exosome |
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