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Improvement of short tandem repeat analysis of samples highly contaminated by humic acid |
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| Authors: | Seung Bum Seo Hong Xuan Jin Hye Young Lee Jianye Ge Jonathan L King Sung Hee Lyoo Dong Hoon Shin Soong Deok Lee |
| Institution: | 1. Department of Forensic Medicine, College of Medicine, Seoul National University, Seoul 110-799, South Korea;2. Department of Forensic and Investigative Genetics, Institute of Applied Genetics, University of North Texas Health Science Center, Fort Worth, TX 76107, USA;3. Institute of Forensic Medicine, College of Medicine, Seoul National University, Seoul 110-799, South Korea;4. Department of Anatomy, College of Medicine, Seoul National University, Seoul 110-799, South Korea |
| Abstract: | We investigated several methods for obtaining successful short tandem repeat (STR) results from high-humic acid (HA)-content samples. DNA purification efficiency was tested for QIAquick® PCR Purification, QIAamp® DNA Investigator and Prepfiler? Forensic DNA Extraction kits. HA-removal capacity of Inhibitor Remover and InhibitEX® Tablet was tested. Experiments on overcoming HA effects on STR amplification were conducted using an AmpliTaq Gold® DNA Polymerase and a TaKaRa Ex Taq? Hot Start Version (Ex Taq HS) with BSA addition. QIAquick kit was most efficient in HA removal and Ex Taq HS showed high resistance to HA. Increasing the amounts of Taq polymerases and BSA addition were shown to be efficient in overcoming PCR inhibition, but BSA addition was superior to the former method. Inhibitor Remover and InhibitEX® Tablet did not positively affect the STR results. This study will help achieve better STR results with high-HA-content samples. |
| Keywords: | Forensic DNA typing DNA recovery Short tandem repeat Inhibitor Humic acid |
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