title: Stability of ultramer as copy number standards in real-time PCR.
abstract: Since commercial copy number standards are not always available for real-time PCR, alternative sources of DNA are used. Unfortunately, stored genomic DNA or PCR amplicon has been shown to be unstable, resulting in variable copy number. More recently, the use of ultramer as copy number standard has been reported. However, there is little information on the stability of ultramer under different storage conditions. Thus the aim of this study was to determine the stability of ultramer as copy number standard under different storage conditions using different mixing methods. We found that ultramer copy number was not affected by storage at either 4°C or -20°C over a period of 30 days. Furthermore, the method of mixing the ultramer did not appear to contribute to variability in results. Irrespective of storage temperature or mixing method, there was less than 5% variance in Ct value over a period of 30 days. A duplicate set of standards costs approximately $0.01. Therefore, the use of ultramer as copy number standards in real-time PCR, is cost effective and convenient.
authors.affiliation: Department of Haematology and Cell Biology, University of the Free State, Bloemfontein, South Africa. viljoencd@ufs.ac.za
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Viljoen
authors.forename: Gaynor G
authors.initials: GG
authors.lastname: Thompson
authors.forename: Sandhya
authors.initials: S
authors.lastname: Sreenivasan
mesh_terms: D003080:Cold Temperature; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D018628:Gene Dosage; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D017422:Sequence Analysis, DNA / Q000592:standards
chemical_list: D004247:DNA