Optimisation and Validation of method for DNA extraction from leaf and woody tissues of <i>Dalbergia latifolia</i>

Abstract

Dalbergia latifolia popularly known as Indian Rosewood, is one of the CITES listed forestry timber species often found illegally logged and marketed. One of the promising techniques for identification and authentication of the same is forensic investigation, which requires DNA with amenable quality and quantity. Here, we describe a modified C-TAB protocol for extracting DNA from different tissues of D. latifolia, and also the effect of sample storage at - 20°C for different period of time on the DNA yield. The DNA was initially isolated from leaf and branch samples on 2nd day after collection and storage, and then extracted at every 20 days interval of sample storage until 200 days, and later at 100 days interval up to 500 days. DNA yield was comparatively more in branch tissues, isolated after the different durations of storage, than in leaf tissues. The quantity and quality of DNA during initial 200 days was better compared to later period of sample storage, though sufficient quantity of DNA with amenable quality was obtained even up to 500 days of storage. However, the DNA band quality for leaf and branch tissues showed smeared pattern depicting DNA damage or shearing, as the storage period increased. Hence, it is better to isolate DNA within lower duration of sample storage, and the method reported herein can be utilised for DNA isolation from woody tissues of D. latifolia for further molecular investigations.