title: Illumina HiSeq 2000 paired end sequencing; mutRNAseq_004366-1A_3_C88BAACXX_lane8
library_construction_protocol: Protocols :  Extraction Protocol: RNA isolation was done according to manufacturer instructions for TRIzol LS, including overnight precipitation step with 1 ul glycogen at -80C. RNA was treated with RNAse free DNAse I (Roche) for 30 min and purified with Agencourt RNAclean XP beads (Beckam Coulter) according to manufacturerâ€™s instructions.  Library Protocol: Total RNA was depleted from rRNA, using an strategy based on Khodor et al, Genes and Development 2011. Briefly, 32 biotinylated oligos annealing Drosophila rRNA were used for retrotranscription of 1-5ug of total RNA, and the RNA-cDNA duplex was pulled-down with streptavidin-conjugated Dynabeads. The removal was performed 2 consecutive times. Then, RNA-seq libraries were prepared using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (protocol B). Barcoded Y-shaped adapters were used for ligation, and standard common PCR primers for Illumina were used for amplification, tipically 14-15 cycles.
attributes.tag: Factor Value[Genotype]
attributes.value: XLOC_004366*