Elfriede G. Van den Eeckhout

Comparison of slab gel electrophoresis and capillary electrophoresis for the detection of the fluorescently labeled polymerase chain reaction products of short tandem repeat fragments

The sizing capability of slab gel electrophoresis for short tandem repeat (STR) fragments was compared to the sizing capability of capillary electrophoresis (CE). Both systems used automated laser fluorescence detection to detect four fluorescent dyes, enabling the use of an internal lane standard within each sample. The STR fragments were amplified using a multiplex polymerase chain reaction (PCR) in which the STR fragments Hum CD-4, Hum TH01, Hum D21S11 and Hum SE33 were amplified simultaneously. The reproducibility of the size calling was determined for both systems. The average standard deviation obtained for the slab gel system was 0.2, which was comparable to the standard deviation of 0.12 obtained for the CE system. The CE system produced results comparable to those obtained on the slab gel system, with a level of precision of +/- 1.0 bp (between instruments).

A strategy for the identification of 2′-deoxynucleoside and 2′--deoxynucleotide adducts using electrospray tandem mass spectrometry

A methodology using electrospray tandem mass spectrometry (ES MS/MS) was developed to identify 2′-deoxynucleotide adducts of bisphenol A diglycidylether (BPADGE). The 2′-deoxynucleotide adducts were formed in vitro and were separated using reversed-phase high-performance liquid chromatography (RP-HPLC). The adducts were investigated by (–) electrospray liquid chromatography-mass spectrometry [(–) ES LC-MS]. (–) ES LC-MS/MS allowed the differentiation between 5′-phosphate- and base-alkylated adducts. More detailed structural information on the heterocyclic moiety was obtained by examining the (+) ES LC-MS/MS data and by comparison with the (+) ES LC-MS/MS spectra of the corresponding 2′-deoxynucleoside adducts.