Workflow from Scientific Research

TEMPOmap experimental workflow. After 5-EU-labeled cells are prepared, a set of tri-probes (splint, primer and padlock) are conjugated or hybridized to cellular mRNAs (Extended Data Fig. 1b,i for more details), resulting in the enzymatic replication of each padlock sequence into cDNA amplicons. The amplicons are anchored in situ via a functionalized acrylic group (blue) to a hydrogel mesh to create a DNA-gel hybrid (blue wavy lines). The five-base barcode on each amplicon is read out by six rounds of SEDAL. Thus, multiplexed RNA quantification reveals gene expression in nascent subcellular locations.

DOI