GSM5386226: MDA-MB-468 - c15ROW35; Homo sapiens; RNA-Seq
Single cell RNA libraries were prepared by Fluidigm C1 Single-Cell Autoprep System. We started scPolyA-seq library preparation by cell lysis. Then we added primers containing polyT and cell barcode for labeling the 3' end of transcripts. Full-length cDNA was synthesized by template-switching reverse transcription, amplification and tagmentation with Tn5 transposases. Nextera XT DNA Library Preparation Kit (Ilumina) was applied to construct scPolyA-seq library. DNA fragments at the 3' end of the cDNA were captured by targeted PCR, and sequencing indexes were added for amplification. The quality of library was examined by Qubit 3.0 Fluorometer and Agilent 2100 Bioanalyzer. Libraries separated by columns were pooled together and sequenced on Illumina HiSeq2500 to obtain 150-bp paired-end reads.