|
| Status |
Public on Jul 07, 2018 |
| Title |
Het_monosome_3 |
| Sample type |
RNA |
| |
|
| Source name |
Gastrocnemius_Het_monosome
|
| Organism |
Mus musculus |
| Characteristics |
genotype: het
tissue: Gastrocnemius
rna source: monosome
|
| Treatment protocol |
NA
|
| Growth protocol |
Mice were housed in standard housing units, fed ab libitum with rodent laboratory chow. Mice were sacrificed by cervical dislocation, and skeletal muscle was immediately removed, cleaned of any excess fat or connective tissue, and flash frozen in liquid nitrogen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Muscle lysate was layered on a 15%–50% linear sucrose gradient and centrifuged at 37,000 rpm for 170 min. The sucrose gradient was fractionated and UV absorption at 260 nm was recorded. RNA was precipitated by adding 2 volumes of 100% ethanol, redissolved in RNase-free water, and treated with phenol-chloroform. Sucrose fractions containing ≥2 polysomes were pooled for subsequent analysis. Total RNA was extracted from muscle using the methods of Caddick et al. (2006).
|
| Label |
Cy3
|
| Label protocol |
Cy3 labeled cRNA was generated using the Agilent Low-Input QuickAmp Kit following the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
600 ng of labeled cRNA was hybridized to Agilent Sureprint G3 Mouse whole-genome microarrays according to manufacturers instructions
|
| Scan protocol |
Agilent SureScan microarray scanner
|
| Data processing |
Microarray data were log2 transformed and normalized using quantile normalisation
|
| |
|
| Submission date |
Jul 06, 2018 |
| Last update date |
Jul 07, 2018 |
| Contact name |
Kim Clarke |
| Organization name |
University of Liverpool
|
| Street address |
Crown Street
|
| City |
Liverpool |
| ZIP/Postal code |
L69 7ZB |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL13912 |
| Series (1) |
| GSE116715 |
mRNA loading on polysomes in skeletal muscle of Eif6 heterozygous mice |
|
