PMC:2041973 / 66952-68171
Annnotations
{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/2041973","sourcedb":"PMC","sourceid":"2041973","source_url":"http://www.ncbi.nlm.nih.gov/pmc/2041973","text":"RNA preparation and array hybridization.\nTotal RNA from cells was processed as follows. Cells were lysed in 1 mL of RNA-bee (Teltest). The RNA was isolated by chloroform extraction of the aqueous phase, followed by isopropanol precipitation as per the manufacturer's instructions. The precipitated RNA was washed in 75% ethanol and eluted with DEPC-treated water. Five ug of RNA was treated with RQ1 DNAase (Promega) according to the manufacturer's instructions. One ug of total RNA for each sample was processed using the Affymetrix GeneChip Whole Transcript Sense Target Labeling Assay (Affymetrix). Ribosomal RNA was reduced with the RiboMinus Kit (Invitrogen). Target material was prepared using commercially available Affymetrix GeneChip WT cDNA Synthesis Kit, WT cDNA Amplification Kit, and WT Terminal Labeling Kit (Affymetrix) as per manufacturer's instructions. Hybridization cocktails containing ∼5 ug of fragmented and labeled DNA target were prepared and applied to GeneChip Human Exon 1.0 ST arrays. Hybridization was performed for 16 hours using the Fluidics 450 station. Arrays were scanned using the Affymetrix 3000 7G scanner and GeneChip Operating Software version 1.4 to produce .CEL intensity files.","divisions":[{"label":"Title","span":{"begin":0,"end":40}}],"tracks":[{"project":"CellFinder","denotations":[{"id":"T157","span":{"begin":375,"end":378},"obj":"CellComponent"},{"id":"T158","span":{"begin":400,"end":406},"obj":"GeneProtein"},{"id":"T159","span":{"begin":987,"end":992},"obj":"Species"},{"id":"T160","span":{"begin":938,"end":941},"obj":"CellComponent"},{"id":"T161","span":{"begin":298,"end":301},"obj":"CellComponent"},{"id":"T162","span":{"begin":139,"end":142},"obj":"CellComponent"},{"id":"T163","span":{"begin":0,"end":3},"obj":"CellComponent"},{"id":"T164","span":{"begin":47,"end":50},"obj":"CellComponent"},{"id":"T165","span":{"begin":479,"end":482},"obj":"CellComponent"},{"id":"T166","span":{"begin":602,"end":615},"obj":"CellComponent"}],"attributes":[{"subj":"T157","pred":"source","obj":"CellFinder"},{"subj":"T158","pred":"source","obj":"CellFinder"},{"subj":"T159","pred":"source","obj":"CellFinder"},{"subj":"T160","pred":"source","obj":"CellFinder"},{"subj":"T161","pred":"source","obj":"CellFinder"},{"subj":"T162","pred":"source","obj":"CellFinder"},{"subj":"T163","pred":"source","obj":"CellFinder"},{"subj":"T164","pred":"source","obj":"CellFinder"},{"subj":"T165","pred":"source","obj":"CellFinder"},{"subj":"T166","pred":"source","obj":"CellFinder"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"CellFinder","color":"#ec93aa","default":true}]}]}}