4.6 Article

MARIS: Method for Analyzing RNA following Intracellular Sorting

Journal

PLOS ONE
Volume 9, Issue 3, Pages -

Publisher

PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0089459

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Funding

  1. Sternlicht Director's Fund Fellowship
  2. Howard Hughes Medical Institute
  3. Harvard Stem Cell Institute
  4. National Institutes of Health [2U01DK07247307, RL1DK081184, 1U01HL10040804]

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Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human beta cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines.

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