GSM2771751: Nanobiopsy Sample_2_9; Homo sapiens; RNA-Seq
Neuron cell bodies and neurites were sampled using the nanobiopsy platform developed by our laboratory. Each nanobiopsy sample was ~50 fL corresponding to ~1% of the cell's total volume. Aspirated RNA samples were processed to cDNA using the ultra low RNA input SMART-Seq system (Takara Bio USA, Mountain View, CA). The cDNA was prepared for each individual aspiration for library preparation.50 ul of 1:5,000 dilution ERCC RNA Spike-In Mix (Ambion) was added to each sample for quality control. cDNA was amplified by KAPA HiFi HotStart ReadyMix PCR Kit using SMART-Seq PCR primers. cDNA was purified by AmPure XP beads. Tagmentation of cDNA for the preparation of RNA-Seq libraries was performed by Nextera XT DNA Library preparation kit (Illumina, San Diego, CA). Multiplexed sequencing adapters were ligated to tagmented cDNA by limited-cycle polymerase chain reaction (10 cycles), according to manufacturer's instructions. The final, amplified libraries were purified by AmPure XP beads (Beckman Coulter Genomics, Danvers, MA), and size-selected in the range of 200-600 bp using the Caliper LabChip XT system (PerkinElmer, Waltham, MA). The final RNA-Seq libraries were quantified using the Agilent Bioanalyzer 2100 High Sensitivity DNA Assay and KAPA Library Quantification Kit, and pooled together in equal concentration for sequencing. The pooled multiplexed libraries were sequenced for 2 × 150 bp paired-end or 2 × 80 bp paired-end reads on Illumina MiSeq (Illumina, San Diego, CA).
GEO Accession
GSM2771751