GSM2650083: 191_outerCC; Bos taurus; RNA-Seq
We extracted RNA from cumulus cells using the guanidinium thiocyanate-phenol chloroform procedure (Trizol reagent, Thermofisher), with the addition of 0.5ul GlycoBlue Coprecipitant (Thermofisher) as a RNA carrier.  For the oocytes, we added 2l of the lysis buffer (20 IU/l RNase inhibitor (Amresco), 0.2% Triton X-100 (Amresco)) to the tube. We eluted the RNA from CCs in 4ml of a solution containing dNTPs and oligo-dT30VN and proceeded with the polyA+ whole transcriptome amplification with the Smart-seq2 protocol for cDNA amplification. For the oocytes, we added 2l of the lysis buffer (20 IU/l RNase inhibitor (Amresco), 0.2% Triton X-100 (Amresco)) to the tube and proceeded with the polyA+ whole transcriptome amplification with the Smart-seq2 protocol for cDNA amplification. The samples were subjected to 16 cycles of PCR amplification. For all samples, we used one ng of amplified cDNA for library preparation using the Nextera XT DNA library preg Kit (Illumina, Inc), as and followed the procedures described in the Smart-seq2 protocol.
GEO Accession
GSM2650083