4.8 Article

Quantifying RNA allelic ratios by microfluidic multiplex PCR and sequencing

Journal

NATURE METHODS
Volume 11, Issue 1, Pages 51-+

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.2736

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Funding

  1. Stanford University School of Medicine
  2. Stanford Graduate Fellowship
  3. US National Institutes of Health [GM102484]
  4. Ellison Medical Foundation
  5. United States-Israel Binational Science Foundation
  6. Edward Mallinckrodt, Jr. Foundation
  7. NATIONAL HUMAN GENOME RESEARCH INSTITUTE [T32HG000044] Funding Source: NIH RePORTER
  8. NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM102484] Funding Source: NIH RePORTER

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We developed a targeted RNA sequencing method that couples microfluidics-based multiplex PCR and deep sequencing (mmPCR-seq) to uniformly and simultaneously amplify up to 960 loci in 48 samples independently of their gene expression levels and to accurately and cost-effectively measure allelic ratios even for low-quantity or low-quality RNA samples. We applied mmPCR-seq to RNA editing and allele-specific expression studies. mmPCR-seq complements RNA-seq for studying allelic variations in the transcriptome.

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