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Collection and Extraction of Saliva DNA for Next Generation Sequencing
Published on: August 27, 2014
Comparison of four DNA extraction methods to extract DNA from cigarette butts collected in Lebanon
Hany Kallassy1, Louis Y El Khoury2, Madona Eid1
1Molecular Biology Laboratory, American University of Science and Technology, Beirut, Lebanon.
Abstract:
Cigarette butts collected from crime scenes represent valuable sources of DNA. However the extraction of the genetic material may deem challenging especially when different contaminants may compromise the integrity, quality, and quantity of DNA obtained. This study aims at comparing four extraction methods (Chelex-100, soaking + Chelex-100, Chelex-100 + PK, and DNA IQ™ System) with the intention of identifying the one with maximal recovery rate and profiling success. DNA was extracted using aforementioned four methods from 70 cigarette butts collected from sites across Lebanon. DNA was quantified by qPCR using TaqMan Quantifiler Kit on an Applied Biosystems 7300 SDS instrument and genotypes were obtained using the PowerPlex® 21 kit on an Applied Biosystems 3130 Genetic Analyser. The findings of this work showed that DNA extraction with Chelex-100 + PK is preferred to the other three methods when seeking both, a high yield and the generation of maximal numbers of full profiles. The Chelex-100 + PK method is simple, cost effective, and therefore suitable for routine cigarette butts case studies.
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