%0 Journal Article
%T Rapid preimplantation genetic screening using a handheld, nanopore-based DNA sequencer - Fertility and Sterility
%A Eric Forman
%A Pallavi Gaur
%A Shan Wei
%A Zachary R. Weiss
%A Zev Williams
%J Fertility and Sterility
%D 2018
%R https://doi.org/10.1016/j.fertnstert.2018.06.014
%X To determine if a handheld, nanopore-based DNA sequencer can be used for rapid preimplantation genetic screening (PGS). Laboratory study. Academic medical center. Amplified genomic DNA from euploid and aneuploid trophectoderm biopsy samples (n=9) that was also tested using traditional next generation sequencing (NGS). Short-read DNA library preparation and nanopore-based sequencing using a hand-held MinION sequencer. Comparison of cytogenetic testing result from NGS and nanopore-based sequencing and the time required for library preparation and sequencing. Multiplexed short-read DNA library preparation was completed in 45 minutes. Sequencing on a single sample was completed within 20 minutes and 5 samples were simultaneously sequenced in under 2 hours. Whole-chromosome aneuploidy screening results obtained from nanopore-based sequencing were identical to those obtained using NGS. Here we report the first application of nanopore-based sequencing for PGS on trophectoderm biopsy samples using a novel rapid multiplxed short-read nanopore sequencing library preparation protocol. Sequencing for aneuploidy screening could be performed on a single sample in 20 minutes and on 5 samples, simultaneously, within 2 hours. Overall, nanopore sequencing is a promising tool to perform rapid PGS onsite, enabling same day testing and embryo transfer, thus obviating the need for complex, large and expensive DNA sequencers or embryo freezing. Cribado gen谷tico preimplantacional r芍pido utilizando la secuenciaci車n manual de ADN basado en nanoporos Determinar si la secuenciaci車n manual de ADN basado en nanoporos puede ser utilizada para el cribado gen谷tico preimplantacional r芍pido. ESTUDIO retrospectivo. Centro m谷dico acad谷mico. Muestras de ADN ampliado procedentes de biopsias de trofoectodermo tanto euploides como aneuploides (n=9), que fueron analizadas seg迆n la t谷cnica de secuenciaci車n masiva cl芍sica (NGS). Preparaci車n de la librer赤a de DNA y secuenciaci車n manual de ADN minION basado en nanosporos. Comparaci車n de los resultados citogen谷ticos obtenidos tras NGS y secuenciaci車n basada en nanosporos y el tiempo empleado para la preparaci車n de la librer赤a y la secuenciaci車n. La preparaci車n de la librer赤a de ADN de lectura corta multiplexada se complet車 en 45 minutos. La secuenciaci車n de una muestra aislada se complet車 en 20 minutos y 5 muestras pudieron ser secuenciadas simult芍neamente por debajo de 2 horas. Los resultados del cribado de aneuploidias de todos los cromosomas obtenidos por la secuenciaci車n manual de ADN basado en nanoporos fueron id谷nticos a aquellos obtenidos
%U https://www.fertstert.org/article/S0015-0282(18)30490-4/fulltext