Ion xpress plus gdna fragment library preparation kit set

Approximately 1 μg of purified WGA DNA was used for library construction by the Ion Xpress Plus gDNA Fragment Library Preparation Kit Set (Life technologies, Carlsbad, CA, USA) following the manufacturer’s instructions. The quantity of library was determined using Qubit dsDNA HS assay kits (Life technologies) with Qubit fluorometers (Life technologies). The template-positive Ion Sphere Particles were generated using Ion PGM Hi-Q Template Kits (Life technologies) with the Ion OneTouch 2 Instrument (Life technologies) and then enriched with the Ion OneTouch ES Instrument (Life technologies). Sequencing was performed on the Ion Torrent PGM Instrument (Life technologies) platform with the Ion PGM Hi-Q Sequencing Kit and Ion 316 chip (Life technologies). The sequencing data analysis was performed by using the cloud-based the Ion Reporter^TM Server System (https://ionreporter.thermofisher.com/ir/).

WGA products of all the biopsied blastocysts were also processed for the NGS analysis in order to evaluate the reliability of rapid aCGH and compare the performances of difference platforms (i.e. rapid aCGH, regular aCGH and NGS). Approximately 1 μg of WGA DNA was used for library construction using Ion Xpress Plus gDNA Fragment Library Preparation Kit Set (Life technologies, California, USA) and following the manufacturer’s instructions. The quantity of library was determined using Qubit dsDNA HS assay kits (Life technologies) with Qubit fluorometer (Life technologies). The template-positive Ion Sphere Particles (ISPs) were generated using Ion PGM Hi-Q Template Kits (Life technologies) with the Ion OneTouch 2 Instrument (Life technologies) and then enriched with the Ion OneTouch ES Instrument (Life technologies). Sequencing was performed on the Ion Torrent PGM Instrument (Life technologies) platform using the Ion PGM Hi-Q Sequencing Kit and Ion 316 chip (Life technologies). PGS analysis for aneuploidy detection was performed by the Ion Reporter Server System (https://ionreporter.thermofisher.com/ir/).

Following the manufacturer’s instructions, WGA was performed using the REPLI-g Single cell kit (QIAGEN, Hilden, Germany). Qubit dsDNA HS assay kits examined the DNA concentrations with a Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Approximately 1 µg of WGA DNA was used for library construction using the Ion Xpress Plus gDNA Fragment Library Preparation Kit Set (Thermo Fisher Scientific). Template positive Ion sphere Particles (ISPs) were generated using Ion 200 Hi-Q Template kits (Thermo Fisher Scientific) with the Ion OneTouch 2 instrument (Thermo Fisher Scientific) and then enriched with the Ion OneTouch ES instrument (Thermo Fisher Scientific). Sequencing was performed on the Ion Proton™ Sequencer (Thermo Fisher Scientific) using an Ion 200 Hi-Q sequencing kit.