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Biopreserv Biobank ; 15(4): 360-365, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28586236

RESUMEN

Whole genome amplification (WGA) has become an invaluable method for preserving limited samples of precious stock material and has been used during the past years as an alternative tool to increase the amount of DNA before library preparation for next-generation sequencing. Myelodysplastic syndromes (MDS) are a group of clonal hematopoietic stem cell disorders characterized by presenting somatic mutations in several myeloid-related genes. In this work, targeted deep sequencing has been performed on four paired fresh DNA and WGA DNA samples from bone marrow of MDS patients, to assess the feasibility of using WGA DNA for detecting somatic mutations. The results of this study highlighted that, in general, the sequencing and alignment statistics of fresh DNA and WGA DNA samples were similar. However, after variant calling and when considering variants detected at all frequencies, there was a high level of discordance between fresh DNA and WGA DNA (overall, a higher number of variants was detected in WGA DNA). After proper filtering, a total of three somatic mutations were detected in the cohort. All somatic mutations detected in fresh DNA were also identified in WGA DNA and validated by whole exome sequencing.


Asunto(s)
Análisis Mutacional de ADN/métodos , ADN/normas , Secuenciación de Nucleótidos de Alto Rendimiento/normas , Mutación/genética , Síndromes Mielodisplásicos/genética , Análisis de Secuencia de ADN/normas , ADN/química , ADN/genética , Análisis Mutacional de ADN/normas , Humanos
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