Genome-wide analysis of mRNA polyadenylation in Cortex/Striatum of CPEB4 modified mice [I]

To test the mRNA polyadenylation status in brain of CPEB4 modified mice, we performed poly(U)-chromatography on RNA purified from a pool of cortex and striatum tissues of WT and CPEB4 KOGT/+ and CPEB4 KO WT, CPEB4 KOGT/+ and CPEB4 KO (n=2) were sacrificed at the age of 6 weeks. The cortex and striatum together were quickly dissected, homogenized and total RNA (INPUT) was extracted. The poly(A) RNA fraction was purified by poly(U)-chromatography. Differential elution with 25% or 90% formamide yielded two fractions of mRNAs enriched for short (WASH) or long (ELUTED) poly(A)-tails, respectively. mRNA fractions were extracted, purified and amplified. cDNA were hybridized in GeneChip Mouse PrimeView arrays and CEL files were generated from DAT files using Affymetrix Command Console software.