Neurofibromatosis Type 1 Gene Mutation Analysis Using Sequence Capture and High-throughput Sequencing

Authors

  • Elina Uusitalo
  • Anna Hammais
  • Elina Palonen
  • Annika Brandt
  • Ville-Veikko Mäkelä
  • Roope Kallionpää
  • Eeva-Mari Jouhilahti
  • Minna Pöyhönen
  • Juhani Soini
  • Juha Peltonen
  • Sirkku Peltonen

DOI:

https://doi.org/10.2340/00015555-1843

Keywords:

mutation analysis, neurofibromatosis type 1, next-generation sequencing, pyrosequencing, saliva DNA, target enrichment

Abstract

Neurofibromatosis type 1 syndrome (NF1) is caused by mutations in the NF1 gene. Availability of new sequencing technology prompted us to search for an alternative method for NF1 mutation analysis. Genomic DNA was isolated from saliva avoiding invasive sampling. The NF1 exons with an additional 50bp of flanking intronic sequences were captured and enriched using the SeqCap EZ Choice Library protocol. The captured DNA was sequenced with the Roche/454 GS Junior system. The mean coverages of the targeted regions were 41x and 74x in 2 separate sets of samples. An NF1 mutation was discovered in 10 out of 16 separate patient samples. Our study provides proof of principle that the sequence capture methodology combined with high-throughput sequencing is applicable to NF1 mutation analysis. Deep intronic mutations may however remain undetectable, and change at the DNA level may not predict the outcome at the mRNA or protein levels.

Downloads

Download data is not yet available.

Downloads

Additional Files

Published

2014-04-07

How to Cite

Uusitalo, E., Hammais, A., Palonen, E., Brandt, A., Mäkelä, V.-V., Kallionpää, R., … Peltonen, S. (2014). Neurofibromatosis Type 1 Gene Mutation Analysis Using Sequence Capture and High-throughput Sequencing. Acta Dermato-Venereologica, 94(6), 663–666. https://doi.org/10.2340/00015555-1843

Issue

Section

Articles