Reliability of SE33 typing by capillary electrophoresis
Introduction
Due to its tremendous variability ACTBP2 (SE33) is a powerful short tandem repeat (STR) for forensic DNA-analysis. The presence of numerous interalleles (#.1, #.2, #.3), however, makes high demands on the resolving capacity of the electrophoresis device used [1], [2], [3], [4], [5]. Capillary electrophoresis (CE) is commonly applied for DNA fragment length determination and sequencing. It shows high-precision and reproducible sizing operation, and therefore CE should fit the demands of such a complex STR system like ACTB2 [6], [7], [8], [9].
We investigated, how precise and reliable the ABI Prism™ 310 Genetic Analyzer determines the length of amplified SE33-fragments under standard conditions with special respect to the capacity of separating 1 bp-differences. Standard deviation of fragment lengths were obtained by repeatedly running an allelic ladder. An attempt was made, to find optimized electrophoretic conditions to increase the separation performance.
Section snippets
PCR-amplification and electrophoresis methods
A ladder of 24 sequenced SE33-alleles (Fig. 1), kindly given by A. Junge, Institute of Legal Medicine, University Bonn, Germany [3], [4], [5], was separated 175 times on a ABI Prism™ 310 Genetic Analyzer (PE Applied Biosystems) in a 5–47 cm×50 μm capillary (PE Applied Biosystems; Part No. 4028399) using performance optimized polymer POP-4™ and the corresponding module GS STR Pop 4 with 26 min separation time. Alleles were sized running the GeneScan™ 2.1 software (ROX350™ size standard; local
Results and discussion
The mean value for the standard deviation (σ) was 0.083 bp (min=0.071 bp; max=0.093 bp; Table 2). 99.62% (=4184) of the 4200 measured SE33 alleles were within the ‘±3σ’-interval. 0.38% (=16) showed slightly larger sizes, but were still within the ‘+4σ’-interval. These were observed within three runs showing a general small shift towards larger calculated allele sizes. The results obtained here are comparable to those of other authors using capillary electrophoresis [6], [7], [9] or the ABI
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A study on short tandem repeat ACTBP2 (SE33) in a Sicilian population sample
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