Quantitative Single-Cell mRNA Analysis in Hydrogel Beads.

Autor: Rakszewska A; Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525, AJ, Nijmegen, The Netherlands., Stolper RJ; Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525, AJ, Nijmegen, The Netherlands., Kolasa AB; Warsaw University of Life Sciences-SGGW, Nowoursynowska 166 ST, 02-787, Warszawa, Poland., Piruska A; Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525, AJ, Nijmegen, The Netherlands., Huck WT; Institute for Molecules and Materials, Radboud University Nijmegen, Heyendaalseweg 135, 6525, AJ, Nijmegen, The Netherlands. w.huck@science.ru.nl.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2016 Jun 01; Vol. 55 (23), pp. 6698-701. Date of Electronic Publication: 2016 Apr 14.
DOI: 10.1002/anie.201601969
Abstrakt: 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.
(© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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