Genomic DNA was extracted from amniocytes or umbilical cord blood cells using a Qiagen DNeasy Blood & Tissue Kit (Qiagen). Genomic DNA (50 ng) was prepared as a template to construct a sequencing library and sequenced using a NextSeq CN500 System (Illumina, San Diego, CA, United States). The sequencing results were subjected to bioinformatics analysis and annotated by the chromosome aneuploidy and gene microdeletion analysis software (Berry, Inc., Beijing, China). The whole experiment process was commissioned by Berry, Inc. The clinical evaluation of results showing CNVs (>100 kb) was based on the aforementioned guidelines.
Nextseq cn500 system
Manufactured by Illumina
Sourced in United States
The NextSeq CN500 system is a high-throughput, benchtop sequencing platform designed for a variety of applications. It utilizes Illumina's proven sequencing-by-synthesis technology to deliver accurate and reliable data. The system's core function is to perform genomic analysis and sequencing.
Automatically generated - may contain errors
3 protocols using nextseq cn500 system
1
Chromosomal Aneuploidy and Microdeletion Analysis
2
Targeted NGS Panel for Solid Tumors
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Targeted NGS with hot expression panel (NGS-H) was conducted. The commercial HANDLE Classic NGS Panel-based library construction (Amoy Diagnostics) was designed to target 36 commonly occurring genes in solid tumors. Determinations of fusion, copy number variation, small insertion or deletion (indel), and single nucleotide variant (Supplemental Table S1) were followed by sequencing using the NextSeq CN500 system (Illumina, San Diego, CA). Rearrangements were analyzed using the AmoyDx NGS data analysis system (ANDAS) (Amoy Diagnostics) as previously described. 18 18. Liu, Y. • Wu, S. • Zhou, L. ... Pitfalls in RET fusion detection using break-apart FISH probes in papillary thyroid carcinoma J Clin Endocrinol Metab. 2021; 106:1129-1138 Crossref Scopus ( 14) PubMed Google Scholar An NTRK fusion was indicated if the output of transcript read counts was ≥10 for the known hotspot fusions and ≥20 copies for novel fusions.
3
Exosomal RNA Extraction and Sequencing
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Total RNA from exosomes was extracted using exoRNeasy Serum/ Plasma Maxi Kit (Qiagen; Valencia, CA, USA) as per manufacturer's instructions. miRNA libraries were constructed using QIAseq miRNA Library Kit (Qiagen) and analyzed using automated DNA analyzer (model Qsep100, BiOptic Inc.; Taiwan). miRNA-seq analysis was performed using NextSeq CN500 system (Illumina; San Diego, CA, USA).
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