GSM1632795: 809_21; Bos taurus; RNA-Seq
Homogenization: A Pro Scientific homogeniser model PRO200 (PRO Scientific Inc., Oxford, CT, USA) was used to homogenise the biopsy in TRIzol® (Life Technologies™, Paisley, UK) with 3 sec pulses 10 times. Clean-up: Total RNA was then cleaned up using the Qiagen RNeasy® Plus Mini kit (Qiagen Ltd) Input total RNA was uniformly 4 µg in a volume of 50 µl nuclease-free H2O for each sample. Indexes from Illumina® TruSeq® v2 A and B kits were allocated to specific samples prior to library construction so that at least two unique and compatible bar codes were in each pool which made provision for a pooling strategy following the Illumina® TruSeq® pooling guidelines. After adapter ligation the ds cDNA fragments were enriched by PCR which created the final sequencing cDNA library. In the PCR enrichment stage of the protocol 10 PCR cycles were used altered from 15 PCR cycles stated in the manufacture’s protocol. Seven equimolar pools of 5 multiplexed cDNA sample libraries were constructed. Indexes from Illumina® TruSeq® v2 kits A and B were allocated to specific samples following the Illumina® TruSeq® pooling guidelines.
GEO Accession
GSM1632795