Microwell-seq
EFO:0030002
A platform for high-throughput single-cell RNA-sequencing. An agarose plate with microwells is used to trap 5-10K individual cells. Barcoded magnetic beads are loaded and trapped into each well. Each single bead is conjugated with 107–108 oligonucleotides, which share the same cellular barcode. Each oligonucleotide consists of a primer sequence, a cell barcode, a unique molecular identifier (UMI), and a poly T tail. After incubation of beads and cells in a soft flow of lysis buffer, beads with captured mRNA are retrieved with a magnet. Beads are collected in a tube in which reverse transcription and template switch steps are performed using the Smart-seq2 protocol (Picelli et al., 2013). Amplified cDNA is fragmented by a customized transposase that carries two identical insertion sequences. The 3′ ends of the transcripts are then enriched during library generation using PCR and sequenced using an Illumina Sequencing Platform.
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