NextSeq 500 paired end sequencing; High-throughput parallel imaging and single-cell transcriptomics of microbial eukaryotic plankton
High-throughput parallel imaging and single-cell transcriptomics of microbial eukaryotic plankton
Cells were collected in exponential phase of the growth concentrated by centrifugation at 8000g for 5 min. Libraries were prepared with Magnatrix 8000+ (Nordiag), an eight-channel robotic workstation capable of running custom made scripts for in-tip magnetic bead separations41. Released mRNAâ€“cDNA hybrids were transferred to the robotic workstation where they underwent second strand synthesis, end repair and in vitro transcription. Following transcription, sequencing adaptors are ligated to the RNA, and another round of cDNA synthesis is performed. Each step in the process is followed by a reaction clean-up step.