Optimized amplification of the polymorphic system COL2A1.
The influence of various amplification parameters on the demonstration of COL2A1 patterns was examined in serial experiments. The combination of 6 optimized parameters (concentration of primers, nucleotides, Taq polymerase, K+, Mg2+, number of cycles) led to an approximately tenfold increase in sensitivity and a decrease in allelic drop-out. In unequal mixtures of DNA from 2 individuals the weakest component was detectable in dilutions down to 1:20. In a small population sample (n = 120) 10 alleles could be demonstrated.
Skowasch
K
K
Institut für Rechtsmedizin, Westfälische Wilhelms-Universität Münster, Federal Republic of Germany.
Brinkmann
B
B
D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D005554:Forensic Medicine / Q000379:methods / Q000592:standards; D005784:Gene Amplification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid