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| Status |
Public on Nov 10, 2009 |
| Title |
Unamplified mRNA abundance in miR-124 and mock-transfected cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
To compare total RNA levels in miR-124 and mock-transfected cells (Figure S3), 5-10 ug of total RNA from miR-124-transfected cells or mock-transfected cells or universal reference RNA (Stratagene Cat# 740000) was reverse transcribed with Superscript III (Invitrogen Cat# 18080085) in the presence of aminoallul-dUTP 5-(3-aminoallyl)-dUTP (Ambion Cat# AM8439) and natural dNTPs (GE Healthsciences Cat# US77212) with 10 ug of N9 primer (Invitrogen). Subsequently, amino-allyl-containing cDNAs from miR-124 and mock-transfected cells were covalently linked to Cy5 NHS-monoesters, and universal reference cDNA was covalently linked to Cy3 NHS-monoesters (GE HealthSciences Cat# RPN5661). Cy5- and Cy3-labeled cDNAs were mixed and diluted into 50 ul of solution containing 3x SSC, 25 mM Hepes-NaOH (pH 7.0), 20 ug human Cot-1 DNA (Invitrogen Cat# 15279011), 20 ug of poly(A) RNA (Sigma Cat# P9403), 25 ug of yeast tRNA (Invitrogen Cat# 15401029), and 0.3% SDS. The sample was incubated at 95 C for 2 min, spun at 14,000 rpm for 10 mins in a microcentrifuge, then hybridized at 65 C
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| Overall design |
An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract.
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| Contributor(s) |
Hogan D |
| Citation(s) |
19901979 |
| |
| Submission date |
Oct 30, 2009 |
| Last update date |
Mar 21, 2012 |
| Contact name |
Patrick O Brown |
| E-mail(s) |
pbrown@cmgm.stanford.edu
|
| Organization name |
Stanford university school of medicine
|
| Department |
Department of biochemistry
|
| Street address |
Beckman Center b439
|
| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
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| Platforms (3) |
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| Samples (9)
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| This SubSeries is part of SuperSeries: |
| GSE18889 |
Concordant Regulation of Translation and mRNA Abundance for Hundreds of Targets of a Human microRNA |
|
| Relations |
| BioProject |
PRJNA123849 |
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