GSM4410970: RMB2867; Mus musculus; RNA-Seq
Cell lysis, reverse transcription and cDNA amplification were performed on the C1 Single-Cell Auto Prep IFC, as specified by the manufacturer (Fluidigm Inc. protocol no. 100-5950 B1). The SMARTer Ultra Low RNA Kit (Clontech) was used for cDNA synthesis from the single cells. The resulting cDNA was quantified with a Quant-iT PicoGreen dsDNA Assay Kit (PN P11496; Life Technologies), and quality was checked with High Sensitivity DNA Reagents (PN 5067-4626) according to the manufacturer's instructions (Agilent Technologies). Only cells with high-quality cDNA and concentration higher than 0.05 ng/μl were processed for subsequent library preparation. Illumina NGS library was constructed with Nextera XT DNA Sample Prep kit (Illumina), according to the manufacturer's recommendations (protocol 100-7168 E1). Sequencing was performed on Illumina HiSeq2000 (Illumina) high output mode by multiplexed single-read run with 51 cycles.
GEO Accession
GSM4410970