Developmental validation of a Cannabis sativa STR multiplex system for forensic analysis |
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Authors: | Howard Christopher Gilmore Simon Robertson James Peakall Rod |
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Affiliation: | School of Botany and Zoology, The Australian National University, Canberra, Australia. |
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Abstract: | A developmental validation study based on recommendations of the Scientific Working Group on DNA Analysis Methods (SWGDAM) was conducted on a multiplex system of 10 Cannabis sativa short tandem repeat loci. Amplification of the loci in four multiplex reactions was tested across DNA from dried root, stem, and leaf sources, and DNA from fresh, frozen, and dried leaf tissue with a template DNA range of 10.0-0.01 ng. The loci were amplified and scored consistently for all DNA sources when DNA template was in the range of 10.0-1.0 ng. Some allelic dropout and PCR failure occurred in reactions with lower template DNA amounts. Overall, amplification was best using 10.0 ng of template DNA from dried leaf tissue indicating that this is the optimal source material. Cross species amplification was observed in Humulus lupulus for three loci but there was no allelic overlap. This is the first study following SWGDAM validation guidelines to validate short tandem repeat markers for forensic use in plants. |
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Keywords: | forensic science DNA typing Cannabis sativa short tandem repeat validation SWGDAM ANUCS301 ANUCS302 ANUCS303 ANUCS304 ANUCS305 ANUCS308 ANUCS501 B01‐CANN1 B05‐CANN1 B02‐CANN2 C11‐CANN1 |
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