GSM2845405: Rat 67; Rattus norvegicus; RNA-Seq
Illumina TruSeq RNA Sample Prep Kit v2 (Illumina, Inc.) was used with 100–1000 ng of total RNA for the construction of sequencing libraries. In short, total RNA samples were poly(A)-selected and subsequently reverser transcribed into double-stranded cDNA. The following steps were cDNA fragmentation, end-repair, and polyadenylation before ligation of the TruSeq adapters containing the index for multiplexing. We checked the quality and quantity of the libraries with a Qubit (1.0) fluorometer and the Caliper LabChip® GX (Caliper Life Sciences, Inc.). The product had an average fragment size of ∼260bp. We normalized the libraries to 10 nm in 10 mm Tris-Cl, pH 8.5 with 0.1% Tween 20. Total RNA samples were poly(A)-selected and subsequently reverser transcribed into double-stranded cDNA. The following steps were cDNA fragmentation, end-repair, and polyadenylation before ligation of the TruSeq adapters containing the index for multiplexing. We checked the quality and quantity of the libraries with a Qubit (1.0) fluorometer and the Caliper LabChip® GX (Caliper Life Sciences, Inc.). The product had an average fragment size of ∼260bp. We normalized the libraries to 10 nm in 10 mm Tris-Cl, pH 8.5 with 0.1% Tween 20. The TruSeq SR Cluster kit v3-cBot-HS (Illumina, Inc.) was used for cluster generation. Sequencing was performed on the Illumina HiSeq 2000 single end 100 bp using the TruSeq SBS kit v3-HS (Illumina, Inc.).