The material obtained from each biopsy was amplified using the SurePlex amplification system (Illumina, San Diego, CA, USA) according to the manufacturer’s instructions. PGT was carried out by aCGH using the 24 Sure V3 microarray (Illumina, San Diego, CA, USA) using the protocol described by Fragouli [28 (link), 29 (link)]. The amplified DNA was fluorescently labeled (Fluorescence Labelling System, Illumina). The samples were co-precipitated, denatured, and analyzed by array hybridization (for 16 h). A laser scanner (InnoScan 710, Innopsys, Carbonne, France) was used to excite the fluorophores and read the hybridization images. Hybridization images were stored in TIFF format and analyzed by the BlueFuse Multi-Analysis software (Illumina), using the criteria and algorithms recommended by the manufacturer. With this approach, it was possible to determine the chromosome constitution of each embryo.
Fluorescence labelling system
Manufactured by Illumina
The Fluorescence Labelling System is a lab equipment product designed for fluorescent labeling of biomolecules. It provides the necessary components and tools to enable the attachment of fluorescent dyes to samples, facilitating their detection and analysis using compatible instrumentation.
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Lab products found in correlation
2 protocols using fluorescence labelling system
1
Comprehensive Chromosome Screening of Embryos
2
Microarray-CGH Analysis of Single Cells
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Microarray-CGH analysis was undertaken according to our previously validated protocol using 24Sure Cytochip (Illumina, Cambridge, UK) (see online supplementary tables S5 and S6). Lysis and whole genome amplification of single cells biopsied from embryos were achieved using the SurePlex kit (Illumina). The entirety of this procedure took place according to the manufacturer's instructions. The fluorescence labelling system (Illumina) was used for the labelling of the amplified ethylenediaminetetraacetic acid-Tris (TE) samples and also for labelling a commercially available reference DNA (Illumina). Test TE samples were labelled with Cy3 while the reference 46,XY DNA was labelled with Cy5. Test and reference DNAs’ co-precipitation, their denaturation, array hybridisation and the posthybridisation washes all took place according to protocols provided by the manufacturer. The hybridisation time was 16 h.
A laser scanner (InnoScan 710, Innopsys, Carbonne, France) was used to analyse the microarrays after washing and drying. The resulting images were stored in TIFF format file and examined by the BlueFuse Multi analysis software (Illumina). Chromosome profiles were examined for gain or loss with the use of a 3× SD assessment.
A laser scanner (InnoScan 710, Innopsys, Carbonne, France) was used to analyse the microarrays after washing and drying. The resulting images were stored in TIFF format file and examined by the BlueFuse Multi analysis software (Illumina). Chromosome profiles were examined for gain or loss with the use of a 3× SD assessment.
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