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Quantitative Single‐Cell mRNA Analysis in Hydrogel Beads
Author(s) -
Rakszewska Agata,
Stolper Rosa J.,
Kolasa Anna B.,
Piruska Aigars,
Huck Wilhelm T. S.
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201601969
Subject(s) - rolling circle replication , lysis , fluorescence , messenger rna , bead , dna , chemistry , single cell analysis , nanotechnology , matrix (chemical analysis) , biophysics , microbiology and biotechnology , cell , computational biology , materials science , biology , physics , chromatography , biochemistry , gene , polymerase , composite material , quantum mechanics
Abstract In recent years, technologies capable of analyzing single cells have emerged that are transforming many fields of biological research. Herein we report how DNA‐functionalized hydrogel beads can serve as a matrix to capture mRNA from lysed single cells. mRNA quantification free of pre‐amplification bias is ensured by using padlock probes and rolling circle amplification followed by hybridization with fluorescent probes. The number of transcripts in individual cells is assessed by simply counting fluorescent dots inside gel beads. The method extends the potential of existing techniques and provides a general platform for capturing molecules of interest from single cells.