GSM1210902: Rrp6; Saccharomyces cerevisiae; RNA-Seq
RNA was extracted using standard Guanidium Thiocyanate - Phenol extraction protocols All directional cDNA libraries were prepared using the Epicentre ScriptSeq kit, as described by the manufacturer’s procedures. 250 ng of RNA was used for ScriptSeq cDNA library preparation and 50 base pairs of each cDNA were sequenced on the Illumina GAIIX system. To prepare the total RNA sequencing libraries, the prp40-1, rrp6 and BY4741 were grown in YPD to an OD=0.3 and subsequently shifted to 37°C for two hours. Prior to preparing the RNA seq cDNA libraries, 4 μg of total RNA was treated with turbo DNase I (Ambion) to remove chromosomal DNA. To enrich for mRNAs, rRNAs were depleted using the RiboMinus kit (Invitrogen). After each step, the integrity of the RNA was analyzed on an Agilent Bioanalyzer 2100. Subsequently, 250 ng of mRNA enriched RNA was used for cDNA library preparation and cDNAs were sequenced on the Illumina HiSeq 2000 v3 platform using the 100 bp paired-end protocol according to manufacturer’s procedures. Samples were multiplexed using the Illumina indexing barcodes. The following barcodes were used for preparation of multiplexed libraries: ATCACG SmD1-TAP_1 CGATGT Control_1 TTAGGC SmD1-TAP_2 TGACCA Control_2 CGATGT BY4741 TTAGGC Rrp6 TGACCA Prp40
GEO
GSM1210902