Increased amplification success from forensic samples with locked nucleic acids.
Inadequate sample quantities and qualities can commonly result in poor DNA amplification success rates for forensic case samples. In some instances, modifying the PCR protocol or components may assist profiling by overcoming inhibition, or reducing the threshold required for successful amplification and detection. Incorporation of locked nucleic acids (LNAs) into PCR primers has previously been shown to increase amplification success for a range of non-forensic sample types and applications. To investigate their use in a forensic context, the PCR primers for four commonly used STR loci have been redesigned to include LNA bases. The modified LNA primers provided significantly increased amplification success when compared to standard DNA primers, with both high-quality buccal samples and simulated forensic casework samples. Peak heights increased by as much as 5.75× for the singleplex amplifications. When incorporated into multiplexes, the LNA primers continued to outperform standard DNA primers, with increased ease of optimisation, and increased amplification success. The use of LNAs in PCR primers can greatly assist the profiling of a range of samples, and increase success rates from challenging forensic samples.
Ballantyne
Kaye N
KN
Biology Division, Victoria Police Forensic Services Department, Macleod, Victoria, Australia. k.ballantyne@erasmusmc.nl
van Oorschot
Roland A H
RA
Mitchell
R John
RJ
D001483:Base Sequence; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D053803:Forensic Genetics; D005838:Genotype; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020080:Tandem Repeat Sequences
D017931:DNA Primers; D009841:Oligonucleotides; C477371:locked nucleic acid