Taqman reference probe vic

Copy number assays were performed using the Droplet Digital PCR (ddPCR) system (Bio‐Rad Laboratories, Inc.). The GeneAssist Copy Number Assay Workflow Builder (Thermo Fischer) was used to design TaqMan assays on Chr15, SNHG14 (Hs05375107_cn) with FAM dye. TaqMan Copy Number Reference Assay, human, RNase P with VIC dye was used as a reference assay. A total of 22 μL reaction mix was prepared, containing 3.5 μL of template DNA (20 ng/μL) without restriction digestion, 10 μL of 2× ddPCR supermix for probes (no UDP) (Bio‐Rad Laboratories Inc.), 1 μL each 20× TaqMan target probe (FAM) and 20× TaqMan reference probe (VIC) (Applied Biosystems), and 6.5 μL of RNase‐/DNase‐free water (see the Supporting Information section for details). All reactions were prepared in triplicates with one negative control. The reaction mixtures were partitioned using the QX200 Droplet Generator and then transferred to a 96‐well plate and amplified using the C1000 Touch thermal cycler, following the manufacturer's protocol. The samples were then read using the QX200 Droplet Reader. Data acquisition and analysis were performed using QuantaSoft Version 1.7.4.0917, and the Poisson algorithm was used to determine the concentrations of the targets as copies/μL.

Copy number assays were performed using the Droplet Digital PCR (ddPCR) System (Bio-Rad Laboratories, Inc.). The GeneAssist™ Copy Number Assay Workflow Builder (Thermo Fischer, United States) was used to design TaqMan assays on Chr11, PKNOX2 (Hs03289418_cn), Chr15, SNHG14 (Hs05375107_cn), and Chr21, TSPEAR (Hs02835328_cn) with FAM dye. TaqMan™ Copy Number Reference Assay, human, RNase P with VIC dye was used as a reference assay. A total of 22 µL reaction mix was prepared, containing 3.5 µL of template DNA (20 ng/μL) without restriction digestion, 10 µL of 2X ddPCR supermix for probes (No UDP) (Bio-RAD Laboratories Inc.), 1 µL each 20X TaqMan target probe (FAM) and 20X TaqMan reference probe (VIC) (Applied Biosystems, United States), and 6.5 µL of RNase-/DNase-free water. All reactions were prepared in triplicates with one negative control. The reaction mixtures were partitioned using the QX200 Droplet Generator™ and then transferred to a 96-well plate and amplified using the C1000 Touch thermal cycler, as per the manufacturer’s protocol. Then, the samples were read using the QX200 Droplet Reader™. Data acquisition and analysis were performed using QuantaSoft Version 1.7.4.0917, and the Poisson algorithm was used to determine the concentrations of the targets as copies/μL.