Agilent DNA 1000 Kit Bioanalyzer DNA Analysis Part Number:5067-1504

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Extract genomic DNA from cultured cells using the QIAamp DNA Mini Kit (Qiagen)	Separate STR-PCR products with a slight modification of the standard protocol

Upstream tips
Extract genomic DNA from cultured cells using the QIAamp DNA Mini Kit (Qiagen)
Protocol tips
Separate STR-PCR products with a slight modification of the standard protocol

Publication protocol

Fragment Length Analysis: Amplified alleles in the STR-PCR products were separated on the Agilent 2100 Bioanalyzer with the Agilent DNA 1000 Kit according to the standard protocol with slight modification.17 In essence, the DNA chip was first primed with 9 μL of the gel-dye mixture, then 3 μL of the PCR product was added to each of the 12 sample wells along with 3 μL of the internal marker (Agilent). One microliter of the Agilent DNA 1000 ladder, used as a sizing standard, was loaded into the ladder well with 5 μL of the internal marker. After electrophoresis, the amplified alleles from each sample were analyzed using the manufacturer's software, 2100 Expert, provided with the instrument; the size of each fragment was determined based on the ladder and internal standards. In addition, the STR markers were identified based on their size ranges.15 Fragment length analysis was done with the Bioanalyzer for all samples from 3 independent STR-PCR experiments (1, 2, and 3). In comparison, samples from the first experiment (Exp 1) were sent to Eurofins MWG Operon (http://www.eurofinsgenomics.eu) and analyzed using standard procedures of capillary electrophoresis on the Applied Biosystems 3130xl Genetic Analyzer, as described in the service information document (Eurofins MWG Operon).18 The relevant negative sample was also included in each of the FLA analyses as a control.

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