Technical note: Polyvinylpyrrolidone (PVP) and proteinase-K improve the efficiency of DNA extraction from Japanese larch wood and PCR success rate. (November 2021)

• Record Type: Journal Article
• Authors: Lee, Byeong-Ju; Kim, Sohyeon; Lee, Jei-Wan; Lee, Hyun-Mi; Eo, Soo Hyung
• Is Part Of: Forensic science international. Volume 328(2021)
• Publication Date: 2021-11
• Subjects: Wood DNA isolation -- Molecular forensics of wood -- Wood identification -- Chloroplast DNA -- PCR inhibitors; Medical jurisprudence -- Periodicals; Chemistry, Forensic -- Periodicals; Forensic Medicine -- Periodicals; Médecine légale -- Périodiques; Chimie légale -- Périodiques; Gerechtelijke geneeskunde; Gerechtelijke chemie; Gerechtelijke psychiatrie; Chemistry, Forensic; Medical jurisprudence; Electronic journals; Periodicals; Electronic journals; 614.1
• DOI: 10.1016/j.forsciint.2021.111005
• Languages: English
• ISSNs: 0379-0738

Abstract

Highlights: Illegal timber confuses the timber market and causes depletion of forest resources. DNA barcoding is the promising method to track the species and origin of wood. The method is needed to increase the efficiency of DNA extraction from wood. The crushed samples from Japanese larch wood were used in this study. Polyvinylpyrrolidone and proteinase-K increase the efficiency of wood DNA extraction. Abstract: Illegal distribution of timber disrupts the timber market and depletes forest resources. DNA markers are used to verify the legal distribution of wood. However, it is difficult to obtain the quantity and quality of DNA suitable for genetic analysis because of the physicochemical properties of wood; therefore, an efficient wood DNA extraction method is required. In this study, to prepare an efficient DNA extraction method from Japanese larch ( Larix kaempferi ) wood, we investigated the ability of polyvinylpyrrolidone (PVP) and proteinase-K to improve DNA extraction efficiency and PCR success rate. It was found that the addition of PVP resulted in a significant increase in the DNA concentration of the treatment group compared to that of the control group, while the purity (A260/A280) showed no difference. Moreover, the addition of proteinase-K significantly increased both the DNA concentration and purity of the treatment group compared to those of the control group. Further analysis showed that the PCR success rate of psbC (approximately 350 bp) was higher than 90% in the control, PVP treatment, and proteinase-K treatment groups. However, in the PCR success rate of rbcL (approximately 1.3 kb) was higher in the proteinase-K and PVP treatment groups than in the control group. The addition of PVP and proteinase-K increased the success rate of PCR amplification for long regions by preventing DNA damage caused by phenolic compounds and proteins in the wood. The results of this study can thus develop DNA extraction methods to identify the species and origin of woods.