title: Clarity™ digital PCR system: a novel platform for absolute quantification of nucleic acids.
abstract: In recent years, digital polymerase chain reaction (dPCR) has gained recognition in biomedical research as it provides a platform for precise and accurate quantification of nucleic acids without the need for a standard curve. However, this technology has not yet been widely adopted as compared to real-time quantitative PCR due to its more cumbersome workflow arising from the need to sub-divide a PCR sample into a large number of smaller partitions prior to thermal cycling to achieve zero or at least one copy of the target RNA/DNA per partition. A recently launched platform, the Clarity™ system from JN Medsys, simplifies dPCR workflow through the use of a novel chip-in-a-tube technology for sample partitioning. In this study, the performance of Clarity™ was evaluated through quantification of the single-copy human RNase P gene. The system demonstrated high precision and accuracy and also excellent linearity across a range of over 4 orders of magnitude for the absolute quantification of the target gene. Moreover, consistent DNA copy measurements were also attained using a panel of different probe- and dye-based master mixes, demonstrating the system's compatibility with commercial master mixes. The Clarity™ was then compared to the QX100™ droplet dPCR system from Bio-Rad using a set of DNA reference materials, and the copy number concentrations derived from both systems were found to be closely associated. Collectively, the results showed that Clarity™ is a reliable, robust and flexible platform for next-generation genetic analysis.
authors.affiliation: JN Medsys, 217 Henderson Road #02-08, Singapore, 159555, Singapore.
authors.forename: Huiyu
authors.initials: H
authors.lastname: Low
authors.affiliation: Centre for Biomedical and Life Sciences, Singapore Polytechnic, 500 Dover Road, Singapore, 139651, Singapore.
authors.forename: Shun-Jie
authors.initials: SJ
authors.lastname: Chan
authors.affiliation: JN Medsys, 217 Henderson Road #02-08, Singapore, 159555, Singapore.
authors.forename: Guo-Hao
authors.initials: GH
authors.lastname: Soo
authors.affiliation: JN Medsys, 217 Henderson Road #02-08, Singapore, 159555, Singapore.
authors.forename: Belinda
authors.initials: B
authors.lastname: Ling
authors.affiliation: Centre for Biomedical and Life Sciences, Singapore Polytechnic, 500 Dover Road, Singapore, 139651, Singapore. englee@sp.edu.sg.|Department of Paediatrics, University Children's Medical Institute, National University Hospital, 1E Kent Ridge Road, Singapore, 119228, Singapore. englee@sp.edu.sg.
authors.forename: Eng-Lee
authors.initials: EL
authors.lastname: Tan
mesh_terms: D006801:Humans; D009696:Nucleic Acids / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D043262:Ribonuclease P / Q000235:genetics
chemical_list: D009696:Nucleic Acids; D043262:Ribonuclease P
keywords: Absolute DNA quantification; Clarity; Sample partitioning; Tube-strip digital PCR