|
| Status |
Public on Mar 12, 2014 |
| Title |
5min: Block, Biological Replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Hippocampal cell culture
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague Dawley
age: Embryonic day 18.5
cell type: hippocampal neurons
|
| Treatment protocol |
Treatments were for 5 minutes, 30 minutes and 60 minutes
|
| Growth protocol |
Cultures were grown for 12 days prior to treatments
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction followed by cleancup using Qiagen RNeasy column. Quality control by Agilent Bioanalyser
|
| Label |
33P
|
| Label protocol |
3 ug of total RNA was reverse-transcribed using oligo-dT primers (Invitrogen) and SuperScript II reverse transcriptase (Invitrogen) in the presence of 100 mCi of [33P]dCTP (Amersham Bioscience, Piscataway, NJ)
|
| |
|
| Hybridization protocol |
The nylon filters were prehybridized for 2 h at 60C with 0.5 mg/ml of poly-dA (Invitrogen) and 0.5 mg/ml Cot-1 DNA (Invitrogen) in 10 ml of AlkPhos DIRECT hybridization buffer (Amersham Bioscience) and then hybridized for 17 h at 60C with the denatured probes
|
| Scan protocol |
Filters were scanned using Storm phosphoimager
|
| Description |
5min- #84-Block
|
| Data processing |
Scanned images processed using phosphoimager software
|
| |
|
| Submission date |
Mar 11, 2014 |
| Last update date |
Mar 12, 2014 |
| Contact name |
Philip Lee |
| Organization name |
NIH
|
| Department |
NICHD
|
| Street address |
35 Lincoln Drive
|
| City |
Bethesda |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL1211 |
| Series (2) |
| GSE55778 |
Hippocampal neurons, nylon-2 experiments |
| GSE55781 |
Systematic identification of 3'UTR regulatory elements in activity-dependent mRNA stability in hippocampal neurons |
|
