PCR amplification of repetitive sequences as a possible approach in relative species quantification

• Published in: Meat science Vol. 90; no. 2; pp. 438 - 443
• Main Authors: Ballin, N.Z., Vogensen, F.K., Karlsson, A.H.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.02.2012; Elsevier
• Subjects: Animals; Authentication; Biological and medical sciences; Chickens; detection limit; DNA; DNA - isolation & purification; DNA primers; DNA Primers - genetics; Dogs; Food industries; Fundamental and applied biological sciences. Psychology; genome; Horses; Limit of Detection; LUX primers; Meat; Meat - analysis; Meat and meat product industries; polymerase chain reaction; Rabbits; Real time PCR; Real-Time Polymerase Chain Reaction - methods; Repetitive sequences; Repetitive Sequences, Nucleic Acid; Sheep; Species identification; Species Specificity; Swine; Real time PCR; Species identification; Authentication; Repetitive sequences; LUX primers; Meat; Meat product; Quantitative analysis
• ISSN: 0309-1740; 1873-4138
• DOI: 10.1016/j.meatsci.2011.09.002

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.