GSM2829106: lung / dog; Canis lupus familiaris; RNA-Seq
10ug total RNA was poly-A selected twice using Dynabeads mRNA Purification Kit (Invitrogen, 610.06). Resulting mRNA was DNase treated (Ambion AM1907) and then fragmented using heat. First strand cDNA synthesis was performed using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, 11917-010), supplementing in SuperScript III Reverse Transcriptase (Invitrogen, 18080-093), incorporating SUPERase*In (Ambion, AM2694), and Actinomycin D (USB, 10415). First strand cDNA was cleaned using 1.8x RNAClean XP SPRI beads (Beckman Coulter, A64987). Second-Strand Synthesis was performed replacing dTTP with dUTP, and the resulting double-stranded cDNA was cleaned using a MinElute PCR Purification Kit (Qiagen, 28004). Illumina libraries were constructed by repairing the ends of the cDNA, ligating adapters, and cleaning/size-selecting with 0.7x SPRI. Illumina libraries were treated with USER to excise dUTP, and amplified via PCR using Fusion Master mix with GC buffer (NEB, F532S). Samples were sequenced on HiSeq 2000 machines, to a minimum depth of 35 million reads.
GEO
GSM2829106
GEO Accession
GSM2829106