High-throughput Single-Cell Proteomics and Transcriptomics from the Same Cells with a Nanoliter-Scale Spin-Transfer Approach
Following the transfer of samples into a cellenCHIP 384-well plate containing RT-PCR buffer (cellenCHIP 384 RTready (Cellenion, cat# CEC-5015-8)) with modified barcode oligo dT in each well. The reverse transcription reaction was performed at 42C for 90 minutes to generate the first strand cDNA. Following the first strand cDNA synthesis, all samples from cellenCHIP 384 was pooled into one reaction tube for cDNA amplification to generate the second strand of cDNA. 1 ng of the double strand cDNA was fragmented and tagged with specific primer using cellenTAG reagent and cellenTAG enzymes, followed by another round of amplification to enrich the template library for sequencing. Libraries were sequenced on the NextSeq 500 platform (Illumina) with paired-end sequencing and dual indexing. A total of 26, 8 and 100 cycles were run for Read 1, i7 index and Read 2, respectively.
SUB15685176
not applicable