Identification and Characterization of Splicing Defects by Single-Molecule Real-Time Sequencing Technology (PacBio)

M. Savarese, T. Qureshi, A. Torella, P. Laine, T. Giugliano, P.H. Jonson, M. Johari, L. Paulin, G. Piluso, P. Auvinen, V. Nigro, Bjarne Udd, P. Hackman

Research output: Contribution to journalArticleScientificpeer-review


Although DNA-sequencing is the most effective procedure to achieve a molecular diagnosis in genetic diseases, complementary RNA analyses are often required. Reverse-Transcription polymerase chain reaction (RT-PCR) is still a valuable option when the clinical phenotype and/or available DNA-test results address the diagnosis toward a gene of interest or when the splicing effect of a single variant needs to be assessed. We use Single-Molecule Real-Time sequencing to detect and characterize splicing defects and single nucleotide variants in well-known disease genes (DMD, NF1, TTN). After proper optimization, the procedure could be used in the diagnostic setting, simplifying the workflow of cDNA analysis. © 2020 - IOS Press and the authors. All rights reserved.
Original languageEnglish
JournalJournal of Neuromuscular Diseases
Issue number4
Pages (from-to)477-481
Number of pages5
Publication statusPublished - 2020
MoE publication typeA1 Journal article-refereed

Fields of Science

  • Long-read sequencing
  • PacBio
  • single molecule real time
  • splicing
  • complementary DNA
  • Article
  • Becker muscular dystrophy
  • DMD gene
  • DNA determination
  • DNA sequencing
  • Duchenne muscular dystrophy
  • gene
  • gene identification
  • human
  • human tissue
  • missense mutation
  • muscle biopsy
  • neurofibromatosis type 1
  • NF1 gene
  • priority journal
  • reverse transcription polymerase chain reaction
  • RNA analysis
  • single nucleotide polymorphism
  • SMRT sequencing
  • splicing defect
  • TTN gene
  • workflow
  • 1184 Genetics, developmental biology, physiology

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