title: PCR amplification of repetitive sequences as a possible approach in relative species quantification.
abstract: Both relative and absolute quantifications are possible in species quantification when single copy genomic DNA is used. However, amplification of single copy genomic DNA does not allow a limit of detection as low as one obtained from amplification of repetitive sequences. Amplification of repetitive sequences is therefore frequently used in absolute quantification but problems occur in relative quantification as the number of repetitive sequences is unknown. A promising approach was developed where data from amplification of repetitive sequences were used in relative quantification of species in binary mixtures. PCR LUX primers were designed that amplify repetitive and single copy sequences to establish the species dependent number (constants) (SDC) of amplified repetitive sequences per genome. The SDCs and data from amplification of repetitive sequences were tested for their applicability to relatively quantify the amount of chicken DNA in a binary mixture of chicken DNA and pig DNA. However, the designed PCR primers lack the specificity required for regulatory species control.
authors.affiliation: Department of Food Science, Faculty of Life Sciences, University of Copenhagen, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark. nixb@fvst.dk
authors.forename: N Z
authors.initials: NZ
authors.lastname: Ballin
authors.forename: F K
authors.initials: FK
authors.lastname: Vogensen
authors.forename: A H
authors.initials: AH
authors.lastname: Karlsson
mesh_terms: D000818:Animals; D002645:Chickens; D004247:DNA / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D006736:Horses; D057230:Limit of Detection; D008460:Meat / Q000032:analysis*; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012756:Sheep; D013045:Species Specificity; D013552:Swine
chemical_list: D017931:DNA Primers; D004247:DNA