title: GSM6722455: Single granulosa cell (DRaqL - Smart-seq2) SmSeq2DRaqL11_20_S6_IL04; Mus musculus; RNA-Seq
library_construction_protocol: Cells were isolated with LCM in 6.4 μL of cell lysis buffer (0.6 μL of 5× SuperScript II buffer, 0.1 μL of 100 μM Oligo dT VN, 0.8 μL of dNTP mix [25 mM each], 0.25 μL of 40 U/μL recombinant RNase inhibitor [Takara Bio], 0.5 μL of 100 mM DTT, 0.06 μL of 1 M MgCl2 [Sigma-Aldrich], 2 μL of 5 M Betaine [Sigma-Aldrich], 0.16 μL of 1:500,000 diluted ERCC RNA Spike-In Mix 1, 0.4 μL of 5% SDc, and 1.53 μL of DDW) in the caps of PCR tubes, and spun down into the tubes by brief centrifugation.  The isolated cells were lysed at 72°C for 6 min, followed by addition of 2.8 μL quenching buffer (0.7 μL of Triton X-100, 0.7 μL of 2% BSA, and 1.4 μL of 5× SuperScript II buffer [Thermo Fisher]).  Then, 1.6 μL of a template-switching mixture (0.1 μL of 100 μM N-template-switching oligo, 0.25 μL of SuperScript II, 0.25 μL of SuperScript III, 0.2 μL of recombinant RNase inhibitor, and 0.8 μL of DDW) was added, followed by the cycling RT program for DRaqL-Smart-seq2.  Then, cDNA amplification was performed by adding 15 μ