title: Amplification volume reduction on DNA database samples using FTA™ Classic Cards.
abstract: The DNA forensic community always strives towards improvements in aspects such as sensitivity, robustness, and efficacy balanced with cost efficiency. Therefore our laboratory decided to study the feasibility of PCR amplification volume reduction using DNA entrapped in FTA™ Classic Card and to bring cost savings to the laboratory. There were a few concerns the laboratory needed to address. First, the kinetics of the amplification reaction could be significantly altered. Second, an increase in sensitivity might affect interpretation due to increased stochastic effects even though they were pristine samples. Third, statics might cause FTA punches to jump out of its allocated well into another thus causing sample-to-sample contamination. Fourth, the size of the punches might be too small for visual inspection. Last, there would be a limit to the extent of volume reduction due to evaporation and the possible need of re-injection of samples for capillary electrophoresis. The laboratory had successfully optimized a reduced amplification volume of 10 μL for FTA samples.
authors.affiliation: Health Sciences Authority, Applied Sciences Group, Biology Division, DNA Database Laboratory, 7 Maxwell Road, Singapore 069111, Singapore. wong_hang_yee@hsa.gov.sg
authors.forename: Hang Yee
authors.initials: HY
authors.lastname: Wong
authors.forename: Eng Seng Simon
authors.initials: ES
authors.lastname: Lim
authors.forename: Wai Fun
authors.initials: WF
authors.lastname: Tan-Siew
mesh_terms: D016172:DNA Fingerprinting / Q000295:instrumentation*; D019075:Electrophoresis, Capillary; D005240:Feasibility Studies; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction