embryos were collected in 2-hour intervals and aged to generate animals 0-2, 4-6 and 8-10 hour old.
Extracted molecule
total RNA
Extraction protocol
the dechorionated embryos were first incubated with 0.1mg/ml cycloheximide in PBS for 10 minutes on ice, then homogenized with a motorized plastic pellet pestle in a lysis buffer (20mM Tris-HCl, pH7.4, 140mM Kcl, 5mM MgCl2, 0.5mM DTT, 1% Triton X-100, 0.1mg/ml cycloheximide, 1mg/ml heparin, 50unit/ml RNasin) and incubated for 10 minutes on ice. The nuclei and debris were removed by centrifugation at 12,000x g for 10min at 4oC, and supernatants was loaded onto 20-50% sucrose gradients with the same extraction buffer without Triton x-100.
Label
biotin
Hybridization protocol
Following fragmentation, at least 10 microg of cRNA were hybridized on GeneChip Drosophila Genome 2.0 Array.
Description
Gene expression data from embryos during first 10 hours after egg laying to see the global and transcript-specific translation profile
Data processing
Gene expression measures were computed using the Robust Multichip Average (RMA) method and implemented in the Bioconductor R package.
