GSM4767075: Rat 17 Adipose Quantseq; Rattus norvegicus; RNA-Seq
Tissue samples were homogenized on ice and total RNA was extracted using a Lexogen SPLIT RNA Extraction kit following the manufacturers instructions. Libraries were prepared according to Lexogens instructions accompanying the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina and small RNA sequencing kits. Briefly, the Quantseq kit uses total RNA as input, hence no prior poly(A) enrichment  or rRNA depletion is needed. Library generation starts with oligodT priming containing the  Illumina-specific Read 2 linker sequence.  After first strand synthesis the RNA is removed. Second strand synthesis is initiated by random priming and a DNA  polymerase. The random primer contains the Illumina-specific Read 1  linker sequence. At this step Unique Molecular Identifiers (UMIs)  can be introduced by exchanging the Second Strand Synthesis Mix 1  (SS1) from the standard QuantSeq FWD Kit with UMI Second Strand  Synthesis Mix (USS). No purification is required between first and second strand synthesis.  Second strand synthesis is followed by a magnetic bead-based  purification step rendering the protocol compatible with automation. During the library amplification step sequences required  for cluster generation are introduced. NGS reads are generated towards the poly(A) tail and directly  correspond to the mRNA sequence. To pinpoint the exact 3' end,  Libraries were sequenced on the Illumina HiSeq 3000 following the manufacturer's protocols. RNA-seq, miRNA-seq